Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
  • A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
  • A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings

Definitions

• the present invention relates to the use of an enaminocarbonyl compound in combination with a biological control agent as well as to a preparation method of compositions containing a selected enaminocarbonyl compound and a selected biological control agent, and compositions containing the enaminocarbonyl compound and at least one biological control agent.
• the particular enaminocarbonyl compounds which can be used in combination with at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, optionally in the presence of inoculants have the formula (I)
• R ! represents alkyl, haloalkyl, alkenyl, haloalkenyl, alkynyl, cycloalkyl, cycloalkylalkyl,
• A represents pyrid-2-yl or pyrid-4-yl or represents pyrid-3-yl which is optionally substituted in the 6-position by fluorine, chlorine, bromine, methyl, trifluoromethyl or trifluoromethoxy or represents pyridazin-3-yl which is optionally substituted in the 6-position by chlorine or methyl or represents pyrazin-3-yl or represents 2 -chloropyr azin- 5 -y 1 or represents l ,3-thiazol-5-yl which is optionally substituted in the 2-position by chlorine or methyl, or represents a radical pyrimidinyl, pyrazolyl, thiophenyl, oxazolyl, isoxazolyl, 1 ,2,4-oxadiazolyl, isothiazolyl, 1 ,2,4-triazolyl or 1,2,5-thiadiazolyl which is optionally substituted by fluorine, chlorine, bromine, cyano, nitro, Ci-
• X represents halogen, alkyl or haloalkyl
• Y represents halogen, alkyl, haloalkyl, haloalkoxy, azido or cyano.
• Pre erred enaminocarbonyl compounds of formula (I) are compounds of the formula (IA)
• R is methyl, cyclopropyl or 2,2-difluoroethyl
• the compounds of formula (I) in combination with at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, optionally in the presence of inoculants, are suitable for reducing overall damage of plants and plant parts as well as losses in harvested fruits or vegetables caused by insects, mites, nematodes and phytopathogens.
• the compound of formula (1-3) is particularly preferred to be used or employed according to the invention, thus, also if not mentioned explicitly, the naming of a compound (I-l), (1-2) or (1-3) always implies that the compound of formula (1-3) is preferred.
• the biological control agent may be employed or used in any physiologic state such as active or dormant.
• Dormant yeast e.g. may be supplied for example frozen, dried, or lyophilized.
• a first aspect of the present invention refers to a combination comprising an enaminocarbonyl compound of formula (IA):
• (1-1) (1-2) (1-3) and at least one biological control agent selected from bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally an inoculant.
• This combination is suitable for reducing overall damage of plants and plant parts as well as losses in harvested fruits or vegetables caused by insects, mites, nematodes and phytopathogens.
• One preferred embodiment refers to a combination according to the invention wherein the biological control agent is selected from the group consisting of bacteria, fungi, yeasts or viruses.
• Another preferred embodiment refers to a combination according to the invention, wherein the inoculant is selected from a group consisting of bacteria o the genus Rhizobium leguminosarum, Rhizobium tropici, Rhizobium loti, Rhizobium trifolii, Rhizobium meliloti, Rhizobium fredii, Azorhizohium caulinodans, Pseudomonas, Azospirillum, Azotobacter, Streptomyces, Burkholdia, Agrobacterium, Endo-, Ecto-, and Vesicular-Arbuscular Mycorhizza,
• Yet another preferred embodiment refers to a combination according to the invention, wherein the biological control agent is selected from a group consisting of Metschnikowia fructicola, Bacillus subtilis, Bacillus thuringiensis, Paecilomyces lilacinu, Bacillus amyloliquefaciens, Cydia pomonella Granulosis virus, and Metarhizium anisopliae.
• the biological control agent is selected from a group consisting of Metschnikowia fructicola, Bacillus subtilis, Bacillus thuringiensis, Paecilomyces lilacinu, Bacillus amyloliquefaciens, Cydia pomonella Granulosis virus, and Metarhizium anisopliae.
• another aspect refers to the use of an enaminocarbonyl compound (1-1), (1-2) or (1-3) in combination with at least one biological control agent selected from bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, optionally in the presence of inoculants, for reducing overall damage of plants and plant parts as well as losses in harvested fruits or vegetables caused by insects, mites, nematodes and phytopathogens.
• the plant part is a seed or a plant emerging from the seed, wherein the seed is from a conventional or a transgenic plant.
• Yet another preferred embodiment refers to said use wherein the plant is a horticultural crop selected from carrots, pumpkin, squash, zucchini, potato, sweet corn, onions, ornamentals, medicinal herbs, culinary herbs, tomatoes, spinach, pepper, melon, lettuce, cucumber, celery, beets, cabbage, cauliflower, broccoli, Brussels sprouts, turnip cabbage, kale, radish, rutabaga, turnip, asparagus, bean, pea, apples, raspberry, strawberry, banana, mango, grapes, peaches, pears, guava, pineapple, pomegranate, garlic, capsicum, chili, radish, star fruit, tapioca, walnuts, lemon, mandarin, mangold, mushroom, olive, orange, papaya, paprika, passion fruit, peanuts, pecan nuts, prune, pistachio nuts, persimmon, pamplemouse (grapefruit), eggplant, endive, cranberry, gooseberry, hazel nuts, kiwifruit, almonds, amarant
• a third aspect refers to a method for reducing overall damage of plants and plant parts as well as losses in harvested fruits or vegetables caused by insects mites, nematodes and/or phytopathogens comprising the step of simultaneously or sequentially applying a compound (1-1), (1-2) or (1-3) and at least one biological control agent selected from bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally at least an inoculant on the plant, plant parts, harvested fruits or vegetables.
• One preferred embodiment refers to said method, wherein the plant part is a seed or a plant emerging from the seed.
• the seed can be from a conventional or a transgenic plant.
• Another preferred embodiment refers to said method, wherein the plant is a horticultural crop selected from carrots, pumpkin, squash, zucchini, potato, sweet corn, onions, ornamentals, medicinal herbs, culinary herbs, tomatoes, spinach, pepper, melon, lettuce, cucumber, celery, beets, cabbage, cauliflower, broccoli, Brussels sprouts, kohlrabi, kale, radish, rutabaga, turnip, asparagus, bean, pea, apples, raspberry, strawberry, banana, mango, grapes, peaches, pears, guava, pineapple, pomegranate, garlic, capsicum, chili, radish, star fruit, tapioca, walnuts, lemon, mandarin, mangold, mushroom, olive, orange, papaya, paprika, passion fruit, peanuts, pecan nuts, prune, pistachio nuts, persimmon, pamplemouse (grapefruit), eggplant, endive, cranberry, gooseberry, hazel nuts, kiwifruit, almonds, amarant
• Another aspect refers to a formulation comprising a compound of formula (IA) as defined in claim 1 and at least one biological control agent selected from bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes as described herein.
• a biological control agent selected from bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes as described herein.
• the invention is further directed to the preparation of a composition containing a compound (I-
• At least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally an inoculant, for reducing overall damage of plants and plant parts as well as losses in harvested fruits or vegetables caused by insects, mites, nematodes and phytopathogens.
• the invention is also directed to the use of a compound (1-1), (1-2) or (1-3) and at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogemc nematodes, and optionally at least an inoculant, for the treatment of seeds or a plant emerging from the seed.
• a biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogemc nematodes, and optionally at least an inoculant, for the treatment of seeds or a plant emerging from the seed.
• the invention is directed to a method for protecting seeds comprising the step of simultaneously or sequentially applying a compound (1-1), (1-2) or (1-3) and at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogemc nematodes, and optionally at least an inoculant, on a seed or a plant emerging from the seed.
• a biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogemc nematodes, and optionally at least an inoculant, on a seed or a plant emerging from the seed.
• the method is further called "seed treatment”.
• the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally an inoculant may be applied in any desired manner, such as in the form of a seed coating, soil drench, and/or directly in-furrow and/or as a foliar spray and applied either pre-emergence, post- emergence or both.
• the composition can be applied to the seed, the plant or to harvested fruits and vegetables or to the soil wherein the plant is growing or wherein it is desired to grow.
• the yield of plants treated according to the invention is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% higher than the yield of untreated plants when grown under the same environmental conditions.
• the use or the method to use a compound (1-1), (1-2) or (1-3) and at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes simultaneously or sequentially includes the following application methods, namely both above mentioned components may be formulated into a single, stable composition with an agriculturally acceptable shelf life (so called "solo-formulation"), or being combined before or at the time of use (so called “combined-formulations”),
• the expression “combination” stands for the various combinations of the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore- orming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant, in a solo-formulation, in a single "ready-mix” form, in a combined spray combination composed from solo-formulations, such as a "tank-mix”, and especially in a combined use of the single active ingredients when applied in a sequential manner, i.e. one after the other within a reasonably short period, such as a few hours or days, e.g. 2 hours to 7 days.
• the order of applying compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore- forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally the inoculant, is not essential for working the present invention. Accordingly, the term “combination” also encompasses the presence of the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes on or in a plant to be treated or its surrounding, habitat or storage space, e.g.
• the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes to a plant its surrounding, habitat or storage space.
• the compounds (I-l), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally an inoculant are employed or used in a sequential manner, it is preferred to treat the plants or plant parts (which includes seeds and plants emerging from the seed), harvested fruits and vegetables according to the following method: Firstly applying the compound (I-l), (1-2) or (1-3) on the plant or plant parts, and secondly applying the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally the inoculant, to the same plant or plant parts.
• the time periods between the first and the second application within a (crop) growing cycle may vary and depend on the effect to be achieved.
• the first application is done to prevent an infestation of the plant or plant parts with insects, mites, nematodes and/or phytopathogens (this is particularly the case when treating seeds) or to combat the infestation with insects, mites, nematodes and/or phytopathogens (this is particularly the case when treating plants and plant parts)
• the second application is done to prevent or control the infestation with insects, mites, nematodes and/or phytopathogens.
• Control in this context means that the biological control agent is not able to fully exterminate the pests or phytopathogenic fungi but is able to keep the infestation on an acceptable level.
• the treatment of plants or plant parts (which includes seeds and plants emerging from the seed), harvested fruits and vegetables with the compound (I-l), (1-2) or (1-3), preferably compound (1-3), and the biological control agent selected from bacteria, in particular spore - forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant, is carried out directly or by action on their surroundings, habitat or storage space using customary treatment methods, for example dipping, spraying, atomizing, irrigating, evaporating, dusting, fogging, broadcasting, foaming, painting, spreading-on, watering (drenching), drip irrigating.
• customary treatment methods for example dipping, spraying, atomizing, irrigating, evaporating, dusting, fogging, broadcasting, foaming, painting, spreading-on, watering (drenching), drip irrigating.
• spore-forming bacteria comprise spores and other inactive forms of said organisms which can yield in active organisms.
• said organisms are comprised in form of spores in a formulation, e.g., a solo- or combined-formulation.
• nematode comprises eggs, larvae, juvenile and mature forms of said organism.
• said organisms are comprised in form of eggs, larvae, juvenile or mature form in a formulation, e.g., a solo- or combined-formulation.
• a solo- or combined-formulation is the formulation which is applied to the plants to be treated (e.g., in a greenhouse, on a field, in a wood), e.g., a tank formulation comprising the biological control agent in accordance with the present invention and a compound (1-1), (1-2) or (1-3) or a liquid or solid formulation comprising said biological control agent which is applied prior, after or in parallel with a compound (1-1), (1-2) or (1-3) to a plant to be treated.
• a tank formulation comprising the biological control agent in accordance with the present invention and a compound (1-1), (1-2) or (1-3) or a liquid or solid formulation comprising said biological control agent which is applied prior, after or in parallel with a compound (1-1), (1-2) or (1-3) to a plant to be treated.
• plant to be treated encompasses every part of a plant including its root system and the material - e.g., soil or nutrition medium - which is in a radius of at least 10 cm, 20 cm, 30 cm around the caulis or bole of a plant to be treated or which is at least 10 cm, 20 cm, 30 cm around the root system of said plant to be treated, respectively.
• material - e.g., soil or nutrition medium - which is in a radius of at least 10 cm, 20 cm, 30 cm around the caulis or bole of a plant to be treated or which is at least 10 cm, 20 cm, 30 cm around the root system of said plant to be treated, respectively.
• the treatment can be carried out by applying the compound (1-1), (1-2) or (1-3), preferably compound (1-3), and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant, as a solution, a powder (for dry seed treatment), a water-soluble powder (for slurry seed treatment), or by in crusting, by coating with one or more layers containing the compound (I- 1), (1-2) or (1-3), preferably compound (1-3), and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant.
• the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the in
• a compound (1-1), (1-2) or (1-3) and at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally an inoculant, as a combination is advantageous.
• bacteria in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally an inoculant, as a combination.
• the broadening of the activity spectrum to other agricultural pests i.e. insects, acari, nematodes, and phytopathogens
• other agricultural pests i.e. insects, acari, nematodes, and phytopathogens
• resistant strains of such agricultural pests and/or plant diseases can be achieved.
• the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes can be used in a lower application rate and still achieve the sufficient control of the agricultural pests and/or plant diseases. This is particularly visible if application rates for the before mentioned compounds or biological control agents are used where the individual compounds or biological control agents show no or virtually no activity.
• the invention can also result in an - Si - advantageous behavior during formulation or during use, for example during grinding, sieving, emulsifying, dissolving or dispensing; improved storage stability and light stability, advantageous residue formation, improved toxicological or ecotoxicological behaviour, improved properties of the plant, for example better growth, increased harvest yields, a better developed root system, a larger leaf area, greener leaves, stronger shoots, less seed required, lower phytotoxicity, mobilization of the defense system of the plant, good compatibility with plants.
• an enhanced systemic action of the compound (1-1), (1-2) or (1-3) or the biological control agent selected from bacteria, in particular spore- forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes is higher and/or a persistency of the fungicidal, insecticidal, acaricidal and/or nematicidal action is expected.
• a compound (1-1), (1-2) or (1-3) and at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally an inoculant, as a combination is particularly suitable for treating seed.
• a large part of the damage to crop plants caused by harmful agricultural pests and/or plant diseases is triggered by an infection of the seed during storage or after sowing as well as during and after germination of the plant. This phase is particularly critical since the roots and shoots of the growing plant are particularly sensitive, and even small damage may result in a weak plant (unhealthy plant), reduced yield and even in the death of the plant.
• the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes and optionally an inoculant can be employed or used according to the invention as a solo- or a combined- formulation.
• Such formulations may include agriculturally suitable auxiliaries, solvents, carriers, surfactants and/or extenders.
• bacteria include spore-forming, root-colonizing bacteria, or bacteria useful as bioinsecticide biofungicide and/or bionematicide. Examples o such bacteria to be used or employed according to the invention are:
• Bacillus cereus in particular spores of Bacillus cereus strain
• Bacillus coagulans Bacillus endoparasiticus
• Bacillus endorhythmos Bacillus azotoformans, (1.10) Bacillus kurstaki, (1.1 1) Bacillus lacticola, (1.12) Bacillus lactimorbus, (1.13) Bacillus lactis, (1.14) Bacillus laterosporus, (1.15) Bacillus lentimorbus, (1.16) Bacillus licheniformis, (1.17) Bacillus medusa, (1.18) Bacillus megaterium, (1.19) Bacillus metiens, (1.20) Bacillus natto, (1.21) Bacillus nigrificans, (1.22)
• Bacillus papillae (1.23) Bacillus pumilus, in particular strain GB34 (ATCC 700814, products known as Yield Shield ® ) and strain QST2808 (NRRL No. B-30087, products known as Sonata QST 2808®), (1.24) Bacillus siamensis, (1.25) Bacillus sphaericus (products known as VectoLexs ® ), (1.26) Bacillus subtilis, in particular strain GB03 (ATCC SD-1397, products known as Kodiak ® ) and strain QST713 (NRRL No. B-21661 , products known as Serenade QST 713 ® ), or
• Bacillus subtilis var. amyloliquefaciens strain FZB24 products known as Taegro ®
• Bacillus thuringiensis in particular Bacillus thuringiensis var. israelensis (products known as VectoBac '® ) or Bacillus thuringiensis subsp. aizawai strain ABTS-1857 (products known as XenTari ® ), or Bacillus thuringiensis subsp. kurstaki strain I I D- 1 (products known as Dipel ® ES) or Bacillus thuringiensis subsp.
• Bacillus (abbreviation: B.) is a genus of rod-shaped, gram-positive bacteria, which can produce endospores under stressful environmental conditions. The single species of this genus differ strongly with respect to their usability in the area of plant protection.
• Bacillus subtilis for example the strains GB03 and QST 713, as well as Bacillus amyloliquefaciens, strain FZB 42, are species with phytopathogenic properties. These bacteria are applied to the soil and/or to the leaves.
• Bacillus thuringiensis with its different subspecies produces endotoxin containing crystals which have high insect pathogenic specifity.
• Bacillus thuringiensis subsp, kurstaki, strain I ID- 1 is used for control of lepidopteran larvae, but without noctuidae.
• Bacillus thuringiensis subsp. aizawai for example the strains SAN 401 I. ABG-6305 and ABG-6346, is effective against different lepidopteran species including also noctuidae.
• Bacillus thuringiensis subsp. tenebrionis for example the strains SAN 418 1 and ABG-6479, protects plants against leaf beetle larvae.
• Bacillus thuringiensis subsp. israelensis for example the strains SAN 402 I and ABG-6164, is applied against larvae of various dipteran pests, e.g. mosquitoes and nematoceres.
• bacteria or mutants thereof that have an insecticidal or plant growth promoting activity are preferred to be used or employed in the present invention, in one embodiment in combination with compound (1-3), optionally in the presence of an inoculant.
• bacteria or mutants thereof that have a fungicidal activity are preferred to be used or employed in the present invention, in one embodiment in combination with compound (1-3), optionally in the presence of an inoculant.
• the bacteria given under the numbers (1.4), (1.5), (1.6), (1.15), (1.16), (1.17), (1.22), (1.23), (1.26), (1.27) and (1.36) to (1.44) are to be used or employed in the present invention, such as in combination with compound (1-3), optionally in the presence of an inoculant.
• These bacteria belong to the class of group 1 bacteria as disclosed in Ash et al., 1991, Lett Appl Microbiology 13, 202-206. The genealogy of group 1 bacteria is also shown in
• Group 1 bacteria can be divided into subgroups depending on the ramification within the group.
• subgroup (1) consists of B. pantothenticus, B, lentus, B. badius, and B. smithi
• subgroup (2) consists of B. azotoformans,, B. firmus, B. circulans, B.benzoevorans, B. simplex, B. marrocanus, B. psychrosaccharolyticus, B. megaterium and B. fastidiosus
• subgroup (3) consists of B. lautus, B. licheniformis, B. subtilis, B. amyloliquifaciens, B. lentimorbus, B. popilliae, B.
• Subgroup (3) can be further divided into subgroup (3 a) consisting of B. lautus, B. licheniformis, B. subtilis, B. amyloliquifaciens, B. lentimorbus, B. popilliae, and B. atrophaeus; subgroup (3b) consisting of B. pumilus, B. cereus, B. thuringiensis, B. medusa, and B. mycoides; and subgroup (3c) consisting of B. coagulans, and B. acidoterrestris.
• the bacteria of subgroup (3), (3a), (3b) or (3c) are to be used or employed in the present invention, such as in combination with compound (1-3), optionally in the presence of an inoculant.
• the bacteria of subgroup (1) are to be used or employed in the present invention, such as in combination with compound (1-3), optionally in the presence of an inoculant.
• the bacteria given under the numbers (1.4), (1.5), (1.23), and (1.26) are to be used or employed in the present invention, such as in combination with compound (1-3), optionally in the presence of an inoculant.
• Bacillus amyloliquefaciens strain IN937a Bacillus amyloliquefaciens strain IN937a
• Bacillus amyloliquefaciens strain FZB42 Bacillus amyloliquefaciens strain FZB42 are used or employed in the present invention, in one embodiment in combination with compound (1-3), optionally in the presence of an inoculant.
• Bacillus cereus strain C CM 1-1562 especially spores are used or employed in the present invention, in one embodiment in combination with compound (1-3), optionally in the presence of an inoculant.
• Bacillus pumilus strain GB34 is used or employed in the present invention, in one embodiment in combination with compound (1-3), optionally in the presence of an inoculant.
• bacteria (1.26a) Bacillus sub ti lis strain GB03 and (1.26b) Bacillus sub ti lis var. amyloliquefaciens strain FZB24 are used or employed in the present invention in one embodiment in combination with compound (1-3), optionally in the presence of an inoculant.
• biological control agents that are summarized under the term “fungi” or "yeasts”.
• BIO 1020 products known as BIO 1020
• Metschnikovia fructicola in particular the strain NRRL Y-30752 (products known as Shemer *)
• Microsphaeropsis ochracea products known as Microx ®
• Muscodor albus in particular strain QST 20799 (products known as QRD300)
• Nomuraea rileyi in particular strains SA86101, GU87401, SR86151, CG128 and VA9101,
• Paecilomyces lilacinus in particular spores of P. lilacinus strain 25 1 (AGAL 89/030550, products known as BioAct ® , cf.
• Penicillium bilaii in particular strain ATCC22348 (products known as JumpStart ® , PB-50, Provide), (2.17) Pichia anomala, in particular strain WRL-076, (2.18) Pseudozyma flocculosa, in particular strain PF-A22 UL (products known as Sporodex ® L), (2.19) Pythium oligandrum DV74 (products known as Polyversum), (2.20) Trichoderma asperellum, in particular strain ICC 012 (products known as Bioten *), (2.21) Trichoderma harzianum, in particular T. harzianum T39 (products known as Trichodex *), and (2.22) Beauveria brongniartii, (2.23) Trichoderma atroviride strain C CM No.
• the fungi and yeasts given under the numbers (2.10), (2.11), and (2.15) are to be used or employed in the present invention, in one embodiment in combination with compound (1-3), optionally in the presence o an inoculant.
• biological control agents that are summarized under the term “protozoas”.
• biological control agents that are summarized under the term “viruses”.
• Gypsy moth (Lymantria dispar) nuclear polyhedrosis virus (NPV), (4.2) Tussock moth ⁇ Lymantriidae) NPV, (4.3) Heliothis NPV, (4.4) Pine sawfly ⁇ Neodiprion) NPV, and (4.5) Codling moth ⁇ Cydia pomonella) granulosis virus (GpGV).
• biological control agents that are summarized under the term "entomopathogenic nematode” are:
• inoculants which may be used or employed according to the invention are bacteria of the genus Rhizobium leguminosarum, Rhizobium tropici, Rhizobium loti, Rhizobium trifolii, Rhizobium meliloti, Rhizobium fredii, Azorhizobium caulinodans, Pseudomonas, Azospirillum,
• the amount of the biological control agent selected from bacteria, in particular spore- orming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic which is used or employed in combination with a compound (1-1), (1-2) or (1-3), preferably with a compound (1-3), optionally in the presence of an inoculant, depends on the final formulation as well as size or type of the plant, plant parts, seeds, harvested fruits and vegetables to be treated.
• the biological control agent to be employed or used according to the invention is present in about 0.0001 % to about 80 % (w/w), preferably in about 5 % to about 75 % (w/w), more preferably about 10 % to about 70 % (w/w) of its solo-formulation or combined- formulation with the compound (1-1), (1-2) or (1-3), and optionally the inoculant.
• bacteria, fungi or yeasts are selected as biological control agent, in particular those who are named as being preferred, namely (2.10), (2.11), and (2.15), it is preferred that these biological control agents or, e.g., their spores are present in a solo-formulation or the combined- formulation in a concentration of at least 10 5 colony forming units (e.g. spores per gram, yeast cells per gram) such as 10 5 - 10 !2 cfu/g, preferably 10 6 - 10 !
• the amount o compound (1-1), (1-2) or (1-3) which is used or employed in combination with the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, optionally in the presence of an inoculant depends on the final formulation as well as size or type of the plant, plant parts, seeds, harvested fruit or vegetable to be treated.
• the compound (1-1), (1-2) or (1-3) to be employed or used according to the invention is present in about 0.1 % to about 80 % (w/w), preferably 1 % to about 60 % (w/w), more preferably about 10 % to about 50 % (w/w) of its solo-formulation or combined- formulation with the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant.
• the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant.
• the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and if present also the inoculant in an synergistic weight ratio.
• the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and if present also the inoculant in an synergistic weight ratio.
• these ratios refer to the ratio within a combined-formulation as well as to the calculative ratio of compound (1-1 ), (1-2) or (1-3) and the biological control agent described herein when both components are applied as mono-formulations to a plant to be treated.
• the ratio can be calculated based on the amount of a compound of formula (I), preferably compound (1-1), (1-2) or (1-3), at the time point of applying said component of a combination according to the invention to a plant or plant part and the amount of a biological control agent shortly prior (e.g., 48 h, 24 h, 12 h, 6 h, 2 h, 1 h) or at the time point of applying said component of a combination according to the invention to a plant or plant part.
• a biological control agent e.g. 48 h, 24 h, 12 h, 6 h, 2 h, 1 h
• a compound (1-1), (1-2) or (1-3) and a biological control agent to a plant or a plant part can take place simultaneously or at different times as long as both components are present on or in the plant after the application(s).
• the biological control agent and a compound of formula (I) preferably compound (1-1), (1-2) or (1-3), are applied at different times and a compound of formula (I) is applied noticeable prior to a biological control agent (e.g.
• the skilled person can determine the concentration of a compound of formula (I), preferably compound (1-1), (1-2) or (1-3), on/in a plant by chemical analysis known in the art, at the time point or shortly before the time point of applying a biological control agent.
• concentration of a biological control agent can be determined using tests as described herein that are also known in the art, at the time point or shortly before the time point of applying a compound of formula (I), preferably compound (1-1), (1-2) or (1-3).
• bacteria/bacteria spore preparation in particular a preparation of spore forming bacteria, is in the range of 500 : 1 to 1 : 1000, preferably in the range of 500:1 to 1 :500, more preferably in a range of 500: 1 to 1 :300. It has to be noted that these ratio ranges refer to a bacteria/bacteria spores preparation (to be combined with a compound (1-1), (1-2) or (1-3) or a preparation of a compound (1-1), (1-2) or (1-3)) of around 10 i0 bacteria/spores per gram preparation of said bacteria/bacteria spores.
• a ratio of 1 : 100 means 1 weight part of a compound (1-1), (1-2) or (1-3) and 100 weight parts of a bacteria/bacteria spore preparation having a bacteria/bacteria spore concentration of 10 10 bacteria/bacteria spores per gram bacteria/bacteria spore preparation are combined (either as a solo formulation, a combined formulation or by separate applications to plants so that the combination is formed on the plant).
• the bacteria/bacteria spore concentration of preparations can be determined by applying methods known in the art and described in this application. To compare weight ratios of a compound (I-
• a biological control agent is Bacillus subtilis, preferably strain GB 03
• the synergistic weight ratio of a compound (1-1), (1-2) or (1-3), preferably compound ⁇ -3, to a preparation of B. subtilis of 10 !0 B. subtilis spores per gram preparation is between 10: 1 and 1 :50 or even between 1 : 1 and 1 :20 such as between 1 : 1 and 1 : 15 or between 1 :3 and 1 : 10.
• a biological control agent is Bacillus thuringiensis, preferably strain ABTS-1857
• the synergistic weight ratio of a compound (1-1), (1-2) or (1-3), preferably compound 1-3, to a preparation B. thuringiensis of 10 !0 B. thuringiensis spores per gram preparation is between 500: 1 and 1 : 100 or even between 500: 1 and 1 :50 such as between 450: 1 and 1 :25 or between 400: 1 and 1 : 15.
• a biological control agent is Bacillus amyloliquefaciens, preferably strain FZB 42
• B. amyloliquefaciens spores per gram preparation is between 100: 1 and 1 :100 or even between 10: 1 and 1 : 100 such as between 1 : 1 and 1 :25, between 1 : 1 and 1 : 10 or between 1 : 1.25 and 1 :5.
• the synergistic weight ratio of a compound (1-1), (1-2) or (1-3) to a fungi or yeast preparation is in the range of 100 : 1 to 1 : 20.000, preferably in the range of 50: 1 to 1 : 10.000 or even in the range of 50: 1 to 1 : 1000. It has to be noted that before mentioned ratios ranges refer to spore/cell preparations of fungi or yeast of around 10 !0 spores (fungi) or cells (yeast) per gram preparation of said fungi or yeast.
• a ratio of 1 : 100 means 1 weight part of a compound (1-1), (1-2) or (1-3) and 100 weight parts of a spore preparation of a fungi or yeast having a spore/cell concentration of 10 !0 spores/cells per gram preparation of said fungi or yeast (without compound (1-1), (1-2) or (1-3) or a preparation of compound (1-1), (1-2) or (i-3)).
• the spore/cell concentration of preparations can be determined by applying methods known in the art and described in this application. To compare weight ratios of a compound (i-1), (1-2) or (1-3) to a fungi or yeast preparation, the skilled person can easily determine the factor between a preparation having a spore/cell concentration different from 10 !0 spores/cells per gram fungi/yeast preparation and a preparation having a spore/ceil concentration of 10 !0 spores/cells per gram preparation to calculate whether a ration of a compound (I-l), (1-2) or (1-3) to a fungi or yeast preparation is within the scope of the above listed ratio ranges.
• a biological control agent is Metschnikowia fr cticola, preferably strain NRRL Y-30752
• the synergistic weight ratio of a compound (I-l), (1-2) or (1-3), preferably compound 1-3, to a preparation of M. fructicola of 10 i0 M. fructicola cells per gram preparation is between 10: 1 and 1 : 100 or even between 1 : 1 and 1 :25 such as between 1 :5 and 1 :25 or between 1 : 10 and 1 :20.
• a biological control agent is Paecilomyces lilacinus, preferably strain 251
• the synergistic weight ratio of a compound (I-l), (1-2) or (1-3), preferably compound 1-3, to a preparation of P. lilacinus of 10 !0 P. lilacinus spores per gram preparation is between 500: 1 and 1 :500 or even between 1 : 1 and 1 :500 such as between 1 :200 and 1 :300 or between 1 :200 and 1 :250.
• a biological control agent is Metarhizium anisopliae, preferably strain F52
• the synergistic weight ratio of a compound (I-l), (1-2) or (1-3), preferably compound 1-3, to a preparation of M. anisopliae of 10 !0 M. anisopliae spores per gram preparation is between 100: 1 and 1 : 100 or even between 10: 1 and 1 :20 such as between 1 : 1 and 1 : 10 or between 1.1 : 1 and 1.1 :3.75.
• a biological control agent is a fungus or yeast and the amount of a fungus spore or yeast cells preparation of 10 !0 spores/cells which is dispersed / ha is at least 50 g/ha, such as 50 - 7500 g/ha, 50 - 5000 g/ha, 50 - 2500 g/ha; at least 250 g/ha (hectare), at least 500 g/ha or at least 1000 g/ha.
• the skilled person can easily calculate the amount of preparation/ha having a fungus spore preparation different from 10 10 spores to match this criterion.
• a biological control agent is Paecilomyces lilacinus, e.g., strain 251, and the amount of a fungus spore preparation (based on a fungus spore preparation of 10 !0 spores / g preparation) which is dispersed / ha is at least 50 g/ha; at least 100 g/ha; at least 1000 g ha; at least 2500 g/ha, such as 2500 - 7500 g/ha, 2500 - 6000 g/ha; or at least 4000 g/ha, such as 4000 - 6000 g/ha, e.g. around 5000 g/ha.
• a biological control agent is Metarhizium anisopliae, e.g., strain F52, and the amount of a fungus spore preparation (based on a fungus spore preparation of 10 !0 spores / g preparation) which is dispersed / ha is at least 50 g/ha, such as 50 - 7500 g/ha, 50 - 1000 g/ha, 50 - 250 g/ha such as 90 - 250 g/ha, or 90 - 225 g/ha.
• a biological control agent is Metschnikowia fructicola, and the amount of a yeast cell preparation (based on a yeast spore preparation of 10 10 spores / g preparation) which is dispersed / ha is at least 50 g/ha, such as 50 - 5000 g/ha, 50 - 2000 g/ha; at least 500 g/ha; at least 1000 g/ha, such as 1000 - 2000 g/ha.
• a biological control agent is a bacterium and the amount of a fungus spore or yeast cells preparation of 10 i0 spores/cells which is dispersed / ha is at least 0.1 g/ha.
• the skilled person can easily calculate the amount of preparation/ha having a bacteria/bacteria spore preparation different from 10 10 spores to match this criterion.
• a biological control agent is B. thuringiensis and the amount of the spore preparation (based on a spore preparation of 10 !0 spores / g preparation) which is dispersed / ha is at least 0.1 g/ha (hectare), such as 0.1 - 5000 g/ha, 0.1 - 10 g/ha, 0.1 - 2 g/ha; at least 0.15 g/ha, such as 0.15 - 1 g/ha, 0.15 - 0.9 g/ha, or even 0.15 - 0.81 g/ha.
• a biological control agent is B. subtilis, e.g., strain GB 03, and the amount of the spore preparation (based on a spore preparation of 10 10 spores /' g preparation) which is dispersed / ha is at least 50 g/ha such as 50 - 5000 g/ha, 50 - 2500 g/ha, 50 - 500 g/ha; at least 100 g/ha, or at least 250 g/ha , such as 100 - 500 g/ha or 250 - 350 g/ha, e.g., around 300 g ha.
• a biological control agent is B.
• amyloliquefaciens and the amount of the spore preparation (based on a spore preparation of 10 !0 spores / g preparation) which is dispersed / ha is at least 500 g/ha, such as 500 - 8000 g/ha, 2500 - 7500 g/ha, or 4000 g/ha to 6000 g/ha such as around 5000 g/ha.
• a biological control agent is a bacterium, fungus or yeast and the amount of a bacteria/bacteria spore, fungus spore or yeast cell preparation of 10 10 spores/cells which is mixed with seeds (e.g., in seed treatment) is at least 0.1 g/kg seeds such as 0.1 g/kg - 10 g/kg, 0.1 g/kg - 1 g/kg.
• seeds e.g., in seed treatment
• the skilled person can easily calculate the amount of preparation/ha having a bacteria/bacteria spore, fungus spore or yeast cell preparation different from 10 10 spores g preparation to match this criterion.
• a biological control agent is a virus and the concentration of the virus after dispersal is at least 50 g/ha such as 50 - 7500 g/ha, 50 - 2500 g/ha, 50 - 1500 g/ha; at least 100 g/ha or at least 150 g/ha.
• a biological control agent is a virus, such as Cydia pomonella granulosis virus and the concentration of the virus after dispersal is at least 50 g ha (hectare) such as 50 - 5000 g/ha, 50 - 2500 g/ha or 50 - 1500 g/ha. in a preferred embodiment, the concentration is around 1000 g/ha such as 900 - 1 100 g/ha. However, also other concentrations are suitable such as such as 100 - 1500 g/ha or 100 - 250 g/ha.
• a biological control agent is a nematode and the concentration of the nematodes is at least 10 6 nematodes/ha, e.g., larval stage nematodes/ha, such as 10 6 - 10 !5 nematodes/ha, e.g., larval stage nematodes/ha, 10 6 - 10 12 nematodes/ha, e.g., larval stage nematodes/ha, at least 10 s nematodes/ha, e.g., larval stage nematodes/ha such as 10 s - 10 !5 nematodes/ha, e.g., larval stage nematodes/ha, 10 8 - 10 !2 nematodes/ha, e.g., larval stage nematodes/ha; or at least 10 9 nematodes/ha, e.g., larval stage nematodes/ha, or at least 10 9 nem
• the ratios between bacteria such as Bacillus amyloliquefaciens, Bacillus suhtilis, Bacillus thuringiensis subsp. aizawai, Bacillus thuringiensis subsp. israeliensis, Bacillus thuringiensis subsp. kurstaki, Bacillus thuringiensis subsp.
• tenebrionis, Bacillus pumilus, Bacillus cereus) and compound (1 -1), (1 -2) or (1-3) in a solo- or combined-formulation or on or in a plant to be treated or its surrounding, habitat or storage space is between 500: 1 to 1 :500, such as between 250: 1 to 1 :250 or even 25: 1 to 1 :25.
• the ratios between fungi such as Metarhizium anisopliae, Paecilomyces lilacinus, Beauveria bassiana, Nomuraea rileyi
• compound (1 -1) in one embodiment of the present invention, is between 125: 1 to 1 : 1250 or even 25: 1 to 1 :250.
• the ratios between nematodes (such as
• Steinernema feltiae and Steinernema carpocapsae) and compound (1 -1), (1-2) or (1-3) in a solo- or combined-formulation or on or in a plant to be treated or its surrounding, habitat or storage space is between 125: 1 to 1 : 125, between 100: 1 to 1 :25 or even 50: 1 to 1 :5.
• the application rate of the biological control agent selected from bacteria, in particular spore- forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes to be employed or used according to the present invention may vary. The skilled person is able to find the appropriate application rate by way of routine experiments.
• biological control agents which are part of the present invention are kept/stored at temperatures below 40°C, for example at temperatures below 37°C such as between 4 °C and 35 °C or between 4 °C and 27 °C.
• biological control agents which are part of the present invention should be applied to a plant or parts thereof.
• biological control agents are part of the present invention are applied to a plant or parts thereof at temperatures below 40°C, for example at temperatures between 15 °C and 32 °C or 20 °C and 27 °C.
• Microorganisms such as fungi or bacteria can be obtained by conventional fermentation processes. The fermentation can be carried out using solid, semi-solid or liquid nutrient media. If spores such as conidia are used, preference is given to solid or semi-solid nutrient media.
• the nutrient media contain the nutrients suitable and known for the cultivation of the respective microorganisms, in particular one or more metabolizable carbon sources or nitrogen sources and mineral salts.
• the fermentation is generally carried out at temperatures between about 3 °C and about 40 °C, preferably between 20 °C and 35 °C.
• a representative fermentation is described in US 5 804 208.
• a fermentation process comprises in general the steps of a) incubating spores such as conidia of a microorganism in or on a nutrition medium (such as agar with further additives such as oatmeal); b) separating spores such as conidia from the nutrition medium after the incubation time, (e.g., by shake off the conidia from the medium, centrifuging, filtrating); and optionally c) preparing an emulsion of said isolated conidia.
• a nutrition medium such as agar with further additives such as oatmeal
• the fungus Metarhizium anisopliae, strain DSM 3884, is known from EP 0 268 177.
• the production of conidia of Metarhizium anisopliae is exemplified in EP 0 794 704 (US 5 804 208).
• a nutrition medium such as oatmeal agar (e.g., composition: 30g of oat flakes and 20g of agar) in a Petri dish was inoculated with, e.g., 3 week old conidia of the Metarhizium anisopliae strain DSM 3884.
• the incubation time to multiply the conidia is, e.g., 3, 4, 5, or 6 days.
• the incubation temperature can be around 7 °C to around 40 °C, e.g. 22° to 25°C.
• the formed conidia was isolated by, e.g., shaking off the conidia.
• the conidia can be stirred with 50ml of water containing 1% of a non-ionic emulsifier such as an emulsifier based on polyoxy-ethylene (20) sorbitan monolaurate (Tween 20®) until a suspension was obtained in which the conidia was present as isolated particles.
• a non-ionic emulsifier such as an emulsifier based on polyoxy-ethylene (20) sorbitan monolaurate (Tween 20®)
• the conidia titer was and can be determined using, e.g., a eubauer chamber.
• the conidia can be stored in closed cases under dry conditions, preferably at temperatures between 0° and 25°C.
• Paecilomyces lilacinus strain 251 was isolated from infected nematode eggs in the Philippines, and correctly described taxonomically in 1974. Optimal laboratory growth of Paecilomyces lilacinus strain 251 occurs at 21-27°C, and does not grow or survive above 36°C (U.S. Environmental Protection Agency, P. lilacinus strain 251 Fact sheet). The following cultivation of Paecilomyces lilacinus is exemplified in Patent Application WO/1994/025579:
• Paecilomyces lilacinus (Thorn) Samson (CBS 143.75), obtained e.g. from the CBS (Central Bureau of Fungal Cultures) in Baarn (The Netherlands), can be maintained on Potato Dextrose Agar (PDA; Ditto laboratories) at 25 °C.
• a conidial suspension was obtained by adding sterilized water (e.g., 5 ml) to a Petri dish containing sporulating mycelium and scraping the surface with a glass rod.
• Liquid cultures were obtained by inoculating conidia of the fungus to minimal salt medium or corn flour medium supplemented with the substrate.
• the minimal salt medium consists of 4.56 g H2P04, 2.77 g KH2 1 IP04 , 0.5 g MgS04 . 71120 and 0.5 g KCI /l, pH 6.0.
• Mycelium can be obtained by centrifuging a, e.g., 6 day old culture of conidia of Paecilomyces lilacinus. For example, cultures can be grown in a shaking water bath for several days at 30°C and 125 strokes per minute. Culture filtrates were obtained by centrifuging cultures for, e.g., 45 min at 9000 g.
• the yeast species Metschnikowia fructicola was isolated from the surface of grape berries (cv. Superior) grown in the central part of Israel. At various stages, individual berries were submersed in sterile distilled water in 100 ml beakers and shaken vigorously for 2 hours on rotary shaker at 120 rpm. Aliquots of 100 ml were removed from the wash liquid and plated on PDA (Potato Dextrose Agar; DIFCO Laboratories, U.S.A.) medium. Following 4-5 days of incubation, yeast colonies were picked randomly according to colony characteristics (color and morphology) and streaked individually on fresh medium to obtain biologically pure cultures. Cultures were further purified by repeated streaking on PDA.
• Metschnikowia fructicola was propagated under aerobic conditions at temperatures ranging from 5°C to 37°C. Optimal growth temperature is between 20°C and 27° C.
• the yeast grows in liquid medium (nutrient broth; Droby et al., 1989) with a neutral pH. The cell density of the yeast generally reached its maximum (stationary stage) growth in 24-48 hours.
• liquid medium nutrient broth; Droby et al., 1989
• the cell density of the yeast generally reached its maximum (stationary stage) growth in 24-48 hours.
• fermentation tanks and industrial growth media were preferred.
• the yeast cells were harvested by centrifugation using conventional laboratory or industrial centrifuges.
• Bacillus thuringiensis were cultured using media and fermentation techniques known in the art (see, for example, Rogoff et al, 1969, J. Invertebrate Path. 14: 122-129; Dulmage et al., 1971 , J.
• a subculture of Bacillus thuringiensis isolates can be used to inoculate the following medium, a peptone, glucose, salts medium: Bacto Peptone 7.5 g/1 Glucose, 1.0 g/'l KH ; PC) 4, 3.4 g/'l K : 1 1 PC) 4, 4.35 g/1 salt solution, 5.0 ml/1 CaCl 2 solution, 5.0 ml/1 salts solution (100 ml) MgS0 4 -7H 2 O, 2.46 g MnS0 4 -H 2 0, 0.04 g ZnS0 4 ⁇ 7H 2 0, 0.28 g FeS0 4 ⁇ 7H 2 O, 0.40 g CaCi 2 solution (100 ml), CaCl 2 2H 2 0, 3.66 g pH 7.2.
• a peptone, glucose, salts medium Bacto Peptone 7.5 g/1 Glucose, 1.0 g/'l KH ; PC) 4, 3.4 g/'l
• the salts solution and CaCl 2 solution were sterilized (e.g., filter-sterilized) and added to the sterilized (e.g., autoclaved and cooked) broth at the time of inoculation. Flasks were incubated at around 30°C on a rotary shaker at 200 rpm for 64 hours.
• the procedure can be readily scaled up to large fermentors by procedures well known in the art.
• the Bacillus thuringiensis spores and crystals, obtained in the fermentation can be isolated by procedures well known in the art. A frequently-used procedure is to subject the harvested fermentation broth to separation techniques, e.g., centrifugation.
• Bacillus subtilis is a naturally occurring bacteria found in soils all over the world.
• Bacillus subtilis strain QST713 was isolated in 1995 by AgraQuest Inc. from soil in a California peach orchard. This product is applied to foliage ( YDEC 2001).
• Bacillus subtilis strain GB03 Bacillus subtilis strain GB03
• Sporulation assay Bacillus subtilis strains were grown in 26 SG medium at 37°C and sporulation was assayed at 24 hours after the end of the exponential phase. The number of spores per ml culture was determined by identifying the number of heat-resistant colony forming units (80°C for 10 min) on I B plates.
• Bacillus subtilis strain Marburg, was grown aerobically in heart infusion broth (Difco Laboratories, Detroit, Mich.) on shaker at about 37°C. From an overnight culture 4 drops were inoculated into 70 ml of pre warmed broth. Growth was measured as optical density at 620 nm. Cells were collected after 3.5 - 4.5 hours in the exponential phase of growth. Centrifugation was carried out at room temperature for 15 min at 7000g (Nanninga, The Journal of Cell Biology. Volume 48, 1971 ⁇ pages 219- 224).
• Bacillus subtilis is active in temperatures between 7°C and 45°C.
• Bacillus amyloliquefaciens strain FZB42 was originally isolated from infested soil in Germany (Krebs et al. (1998) J Plant Dis Prot 105: 181 -197, Chen et al. (2007), Nature Biotechnol. 25(9): 1007-14.). Bacillus amyloliquefaciens strain FZB42 was cultivated in Luria broth (LB - 1% w/v peptone, 0.5% w/v yeast extract, 0.5% w/v NaCl) at 30°C (Fan et al. (2011), Journal of Biotechnology 151 : 303-311).
• the bacteria were grown in Landy medium as described in Koumoutsi et al. (2004), J. Bacteriol. 186: 1084-1096. To prepare surface cultures, the strains were grown in petri dishes containing 1.5% Landy agar for 24 h at 37°C and stored at room temperature prior to MALDI-TOF-MS analysis. Fermentation in liquid media was carried out in flasks at 30°C and 180 rpm in a shaker.
• Cydia pomonella granulosis viruses which are used in the products MADEX (Andermatt Biocontrol) and Granupom (Probis GmbH ) are deposited since 2005 at the German Collection of Microorganisms and Cell Cultures (DSMZ). Isolates used for the production of MADEX (Andermatt Biocontrol), Granupom (Probis GmbH), VIRGO (SipcamS.p.A.) and CARPOVIRUSINE (Arysta LifeScience S.A.S) were all derived from the Mexican isolate originally isolated in 1963 and are not genetically modified. (Virus accession number: GV-0001)
• the identity of the virus produce can be bioanalytically checked against the parent strain by SDS-polyacrylamide-gel electrophoresis of the virus protein sand by Restriction endonuclease analysis of viral DNA.
• CpGV Cydia pomonella Granulosis virus
• CpGV Cydia pomonella Granulovirus
• CpGV derives from the Mexican isolate of CpGV (Tanada, (1964), J. Insect Pathol. 6: 378-
• Nematodes can be reared in liquid culture techniques (see, e.g., US 5,023, 183 which is herewith incorporated by reference in its entirety) and stored, for example, as eggs, larvae in suspension cultures or in clay powder or adult nematodes, e.g.in clay powder. Nematodes can be held in the refrigerator (2-6°C) until use for up to 4 weeks and can be reactivated by suspension in warm water
• the nematode suspension was fixed, checked for the presence of entomopathogenic nematodes using an inverted light microscope, and the number of Stein ernema specimens was determined. Species identification was mostly done at high microscopical magnification using morphological characters of the infective-stage juveniles.
• Entomopathogenic nematodes can be mass-produced by in-vivo or in-vitro methods. Larvae of Galleria mellonella are most commonly used to rear nematodes because f their commercial availability.
• Several researchers e.g., Dutky et al. (1964), J. Insect Pathol. 6: 417-422, or Flanders et al. (1996), J. Econ. Entomol. 89: 373-380
• the nematodes are impregnated onto moist carrier substrates providing substantial interstitial spaces leading to increased gas exchange.
• moist carrier substrates include polyether polyurethane sponge, cedar shavings, peat, vermiculite, etc.
• Nematodes held on the sponge need to be hand-squeezed into water before application, whereas from the other carriers they may be applied directly to the soil as mulch (Neves et al., Neotropical Entomology,
• a bioassay to determine nematode viability is described, e.g., in Simser ((1992) J. of Nematology 24(3):374-378). The Nematode viability was verified by host bioassay. Late instar larvae of the greater wax moth, Galleria mellone, were buried 2.5 cm deep between plants before nematode application (four larvae per replicate), collected after 7 days, placed in petri dishes (9 cm diameter) and held in darkness at ca. 25 °C. Insect mortality (>90%) and subsequent nematode propagation with cadavers demonstrated infectivity of the nematodes. The skilled person is well aware how to adopt this kind of bioassay to different nematode species.
• the preferred application rate of bacteria as biological control agent in particular of spores of the bacteria (1.26a), namely B. subtilis strain GB03, lies in the range of 0,1 to 3 kg/ha.
• the preferred application rate of fungi as biological control agent in particular the fungi Metarhizium anisopliae strain F 52 lies in the range of 0,1 to 3 kg/ha
• yeast Metschnikowia fructicola strain NRRL Y-30752 lies in the range of 0,05 to 8 kg/ha.
• the preferred application rate of protozoa, viruses, and entomopathogenic nematodes as biological control agents lies in the range of 0,5 to 10 kg/ha.
• the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and if applicable also the inoculant on horticultural crops, such as cotton, flax, grapevines, fruit, vegetable, such as Rosaceae sp.
• bacteria in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and if applicable also the inoculant on horticultural crops, such as cotton, flax, grapevines, fruit, vegetable, such as Rosaceae sp.
• pomaceous fruit such as apples and pears, but also stone fruit, such as apricots, cherries, almonds and peaches and soft fruit such as strawberries
• Ribesioidae sp. Juglandaceae sp.
• Betulaceae sp. Anacardiaceae sp., Fagaceae sp., Moraceae sp., Oleaceae sp., Actinidaceae sp., Lauraceae sp., Musaceae sp. (for example banana trees and plantations), Rubiaceae sp.
• Theaceae sp. for example coffee
• Theaceae sp. Sterculiceae sp.
• Rutaceae sp. for example lemons, oranges and grapefruit
• Solanaceae sp. for example tomatoes
• Liliaceae sp. for example lettuce
• Umbelliferae sp. for example lettuce
• Cicurbitaceae sp. for example cucumbers
• a lliaceae sp. for example leek, onions
• Papilionaceae sp. for example peas
• major crop plants such Gramineae sp.
• Poaceae sp. for example sugarcane
• Asteraceae sp. for example sunflowers
• Brassicaceae sp. for example white cabbage, red cabbage, broccoli, cauliflowers, Brussels sprouts, pak choi, turnip cabbage, garden radish, and also oilseed rape, mustard, horseradish and cress
• Fabacae sp. for example beans, peas, peanuts
• Papilionaceae sp. for example soya beans
• Solanaceae sp. for example potatoes
• Chenopodiaceae sp. for example sugar beet, fodder beet, Swiss chard, beetroot
• Horticultural crops particularly includes carrots, pumpkin, squash, zucchini, potato, sweet corn, onions, ornamentals, medicinal herbs, culinary herbs, tomatoes, spinach, pepper, melon, lettuce, cucumber, celery, beets, cabbage, cauliflower, broccoli, Brussels sprouts, turnip cabbage, kale, radish, rutabaga, turnip, asparagus, bean, pea, apples, raspberry, strawberry, banana, mango, grapes, peaches, pears, guava, pineapple, pomegranate, garlic, capsicum, chili, radish, star fruit, tapioca, walnuts, lemon, mandarin, mangold, mushroom, olive, orange, papaya, paprika, passion fruit, peanuts, pecan nuts, prune, pistachio nuts, persimmon, pamplemouse (grapefruit), eggplant, endive, cranberry, gooseberry, hazel nuts, kiwi fruit, almonds, amaranth, apricot, artichoke, avocado, blackberry
• the compound (I-l), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and if applicable also the inoculant on broad acre crops, such as cotton, corn, soybean, cereals, canola, oil seed rape, sugar cane and rice.
• Agrochemical formulations as mentioned herein, in particular solo-formulations and combined-formulations may generally include carrier, which is be understood as meaning a natural or synthetic, organic or inorganic substance which is mixed or combined with the active compounds for better applicability, in particular for application to plants or plant parts or seeds.
• the carrier which may be solid or liquid is generally inert and should be suitable for agricultural use.
• the carrier is a solid carrier.
• the formulations mentioned can be prepared in a manner known per se, for example by mixing the active the compound (I-l), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and
• Suitable additives are all customary formulation auxiliaries, such as, for example, organic solvents, extenders, solvents or diluents, solid carriers and fillers, surfactants (such as adjuvants, emulsifiers, dispersants, protective colloids, wetting agents and tackifiers), dispersants and/or binders or fixatives, preservatives, dyes and pigments, defoamers, inorganic and organic thickeners, water repellents, if appropriate siccatives and UV stabilizers, gibberellins and also water and further processing auxiliaries.
• further processing steps such as, for example, wet grinding, dry grinding or granulation may be required.
• agrochemical formulations as mentioned herein may also generally include suitable solid or liquid carriers are: for example ammonium salts and natural ground minerals, such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as finely divided silica, alumina and natural or synthetic silicates, resins, waxes, solid fertilizers, water, alcohols, especially butanol, organic solvents, mineral oils and vegetable oils, and also derivatives thereof. It is also possible to use mixtures of such carriers.
• suitable solid or liquid carriers are: for example ammonium salts and natural ground minerals, such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as finely divided silica, alumina and natural or synthetic silicates, resins, waxes, solid fertilizers, water, alcohols, especially butanol, organic solvents, mineral oils
• Solid carriers suitable for granules are: for example crushed and fractionated natural minerals, such as calcite, marble, pumice, sepiolite, dolomite, and also synthetic granules of inorganic and organic meals and also granules of organic material, such as sawdust, coconut shells, maize cobs and tobacco stalks.
• agrochemical formulations as mentioned herein may also generally include suitable liquefied gaseous extenders or carriers are liquids which are gaseous at ambient temperature and under atmospheric pressure, for example aerosol propellants, such as butane, propane, nitrogen and carbon dioxide.
• suitable liquefied gaseous extenders or carriers are liquids which are gaseous at ambient temperature and under atmospheric pressure, for example aerosol propellants, such as butane, propane, nitrogen and carbon dioxide.
• agrochemical formulations as mentioned herein may also generally include suitable tackifiers, such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules and latices, such as gum arabic, polyvinyl alcohol, polyvinyl acetate, or else natural phospholipids, such as cephalins and lecithins and synthetic phospholipids can be used in the formulations.
• suitable tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules and latices, such as gum arabic, polyvinyl alcohol, polyvinyl acetate, or else natural phospholipids, such as cephalins and lecithins and synthetic phospholipids can be used in the formulations.
• suitable tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules and latices, such as gum arabic, polyvinyl alcohol, polyvinyl acetate
• agrochemical formulations as mentioned herein may also generally include extender.
• the extender used is water, it is also possible for example, to use organic solvents as auxiliary solvents.
• Suitable liquid solvents are essentially: aromatic compounds, such as xylene, toluene or alkylnaphthalenes, chlorinated aromatic compounds or chlorinated aliphatic hydrocarbons, such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons, such as cyclohexane or paraffins, for example mineral oil fractions, mineral and vegetable oils, alcohols, such as butanol or glycol, and also ethers and esters thereof, ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents, such as dimethylformamide and dimethyl sulphoxide, and also water.
• an agrochemical formulation such as a solo-formulation or a combined- formulation comprises at least one of the following solvents selected from the group consisting of water, ketones, such as acetone, dimethylformamide and dimethyl sulphoxide.
• an agrochemical formulation such as a solo-formulation or a combined-formulation comprises at least one of the following solvents selected from the group consisting of water, ketones, such as acetone, dimethylformamide and dimethyl sulphoxide; and further comprises an emulsifier selected from the group consisting of alkylaryl polyglycolether.
• agrochemical formulations as mentioned herein may also generally include additional further components, such as, for example, surfactants.
• surfactants are emulsifiers, dispersants or wetting agents having ionic or nonionic properties, or mixtures of these surfactants.
• salts of polyacrylic acid salts of lignosulphonic acid, salts of phenolsulphonic acid or naphthalenesulphonic acid, polycondensates of ethylene oxide with fatty alcohols or with fatty acids or with fatty amines, substituted phenols (preferably alkylphenols or arylphenols), salts of sulphosuccinic esters, taurine derivatives (preferably alkyl taurates), phosphoric esters of polyethoxylated alcohols or phenols, fatty esters of polyols, and derivatives of the compounds containing sulphates, sulphonates and phosphates.
• substituted phenols preferably alkylphenols or arylphenols
• salts of sulphosuccinic esters salts of sulphosuccinic esters
• taurine derivatives preferably alkyl taurates
• phosphoric esters of polyethoxylated alcohols or phenols fatty esters of polyol
• a surfactant is required if one of the active compounds and/or one of the inert carriers is insoluble in water and when the application takes place in water.
• the proportion of surfactants is between 5 and 40 per cent by weight of the composition according to the invention.
• colorants such as inorganic pigments, for example iron oxide, titanium oxide, Prussian blue, and organic dyes, such as alizarin dyes, azo dyes and metal phthalocyanine dyes, and trace nutrients, such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc.
• agrochemical formulations as mentioned herein may also generally include other additional components, for example protective colloids, binders, adhesives, thickeners, thixotropic substances, penetrants, stabilizers, sequestering agents, complex formers,
• Agrochemical formulations as mentioned herein can be used in form of aerosols, capsule suspensions, cold-fogging concentrates, warm-fogging concentrates, encapsulated granules, fine granules, flowable concentrates for the treatment of seed, ready-to-use solutions, dustable powders, emulsifiable concentrates, oil-in-water emulsions, water-in-oil emulsions, macrogranules, microgranules, oil-dispersible powders, oil-miscible flowable concentrates, oil-miscible liquids, foams, pastes, pesticide - coated seed, suspension concentrates, suspoemulsion concentrates, soluble concentrates, suspensions, wettable powders, soluble powders, dusts and granules, water-soluble granules or tablets, water-soluble powders for the treatment of seed, wettable powders, natural products and synthetic substances impregnated with active compound, and also microen
• the combinations according to the invention do not only comprise ready-to-use formulations which can be applied with suitable apparatus to the plant or the seed, but also commercial concentrates which have to be diluted with water prior to use.
• composition containing a compound (1-1), (1-2) or (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally an inoculant according to the invention is formulated in a single, stable solution, or emulsion, or suspension.
• the compound (1-1), (1-2) or (1-3) is dissolved in a suitable solvent before the biological control agent is added.
• Suitable solvents are liquid and include petroleum based aromatics, such as xylene, toluene or alkylnaphthalenes, aliphatic hydrocarbons, such as cyclohexane or paraffins, for example petroleum fractions, mineral and vegetable oils, alcohols, such as butanol or glycol as well as their ethers and esters, ketones, such as methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents, such as dimethylformamide and dimethyl sulphoxide.
• the solvent is water.
• the compound (1-1), (1-2) or (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant, are suspended in separate solvents and mixed at the time of application.
• the compound (1-1), (1-2) or (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant are combined in a ready-to-use formulation that exhibits a shelf-life of at least two years.
• the liquid can be sprayed or atomized foliarly or in- furrow at the time of planting the plant.
• the liquid composition can be introduced to the soil before germination of the seed or directly to the soil in contact with the roots by utilizing a variety f techniques including, but not limited to, drip irrigation, sprinklers, soil injection or soil drenching.
• stabilizers and buffers can be added, including alkaline and alkaline earth metal salts and organic acids, such as citric acid and ascorbic acid, inorganic acids, such as hydrochloric acid or sulfuric acid.
• organic acids such as citric acid and ascorbic acid
• inorganic acids such as hydrochloric acid or sulfuric acid.
• Biocides can also be added and can include formaldehydes or formaldehyde-releasing agents and derivatives of benzoic acid, such as p-hydroxybenzoic acid.
• alkane, alkyl, alkene, alkenyl, alkine, alkinyl, aryl when mentioned herein refer to groups containing C1-C20, Ci-Cs or Ci-Ce carbon atoms.
• heteroaryl and other functional groups comprising alkyl, alkenyl, alkinyl or aryl, such as ketones, ethers, amines, etc. may contain C1-C20, Ci-Cs or Ci-Ce carbon atoms.
• combinations which are contrary to the law of nature e.g., C2-aryl
• the term "poly” refers to units of 2- 50000, 2-5000, 2-500, 2-50, 5-500, 50-500, 5-50 subunits.
• the agrochemical formulations or in the use forms of the compound (1-1), (1-2) or (1-3) and the biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and or the inoculant there may be additionally at least one further active compound present.
• active compounds may be insecticides, attractants, sterilants, bactericides, acaricides, nematicides, fungicides, growth regulators, herbicides, fertilizers, safeners and semiochemicals.
• the solid or liquid agrochemical formulations or use forms as mentioned before may further contain functional agents capable of protecting seeds from the harmful effects of selective herbicides such as activated carbon, nutrients (fertilizers), and other agents capable of improving the germination and quality of the products or a combination thereof.
• selective herbicides such as activated carbon, nutrients (fertilizers), and other agents capable of improving the germination and quality of the products or a combination thereof.
• Conventional seeds which can be treated according to the invention are seeds of any plant variety employed in agriculture, in the greenhouse, in forests or in horticulture or in vineyards and include horticultural and broad acre crops.
• this takes the form of seed of cereals (such as wheat, barley, rye, triticale, millet, oats), maize (corn), cotton, soybean, rice, potatoes, sunflowers, beans, coffee, beets (e.g. sugar beets and fodder beets), peanuts, oilseed rape, poppies, olives, coconuts, cacao, sugar cane, tobacco, vegetables (such as tomatoes, cucumbers, onions and lettuce), lawn and ornamental plants (also see below).
• cereals such as wheat, barley, rye, triticale, millet, oats
• maize corn
• cotton soybean
• rice potatoes
• sunflowers sunflowers
• beans coffee
• beets e.g. sugar beets and fodder beets
• Transgenic seeds containing at least one heterologous gene which allows the expression of a polypeptide or protein having insecticidal properties are particularly preferred to be treated according to the invention.
• the heterologous gene in transgenic seed can originate, for example, from microorganisms of the species Bacillus, Rhizobium, Pseudomonas, Serratia, Trichoderma, Clavibacter, Glomus or Gliocladium.
• this heterologous gene is from Bacillus sp., the gene product having activity against the European com borer and/or the Western com rootworm.
• the heterologous gene originates from Bacillus thuringiensis.
• the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant according to the invention can be formulated as a solo-agrochemical formulation or a combined- agrochemical formulation with the aim to be sufficiently stable so that the treatment of the plants, plant parts, seeds, harvested fruits and vegetables does not cause any damage.
• the compound (1-1 ), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant can also be applied directly, that is to say without comprising further components and without having been diluted.
• treatment of the seed may take place at any point in time between harvesting and sowing.
• the seed used is separated from the plant and freed from cobs, shells, stalks, coats, hairs or the flesh of the fruits.
• seed which has been harvested, cleaned and dried to a moisture content of less than 15 % by weight.
• seed which, after drying, has been treated for example, with water and then dried again.
• bacteria in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes
• Agrochemical formulations for treating seeds are solutions, emulsions, suspensions, powders, foams, slurries or other coating materials for seed, and also ULV formulations (cf. US 4,272,417, US 4,245,432, US 4,808,430, US 5,876,739, US 2003/0176428, WO 2002/080675, WO 2002/028186).
• Such seed dressing formulations are prepared in a known manner by mixing the compound (I- 1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-formin bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant with customary additives, such as, for example, customary extenders and also sol vents or diluents, colorants, wetting agents, dispersants, emulsifiers, defoamers, preservatives, secondary thickeners, adhesives, gibber ellins and water as well.
• customary additives such as, for example, customary extenders and also sol vents or diluents, colorants, wetting agents, dispersants, emulsifiers, defoamers, preservatives, secondary thickeners, adhesives, gibber ellins and water as well.
• Suitable colorants that may be present in the seed dressing formulations include all colorants customary for such purposes. Use may be made both of pigments, of sparing solubility in water, and of dyes, which are soluble in water. Examples that may be mentioned include the colorants known under the designations Rhodamine B, C.I. Pigment Red 1 12, and C.I. Solvent Red 1.
• Suitable wetting agents that may be present in the seed dressing formulations include all substances which promote wetting and are customary in the formulation of active agrochemical substances. With preference it is possible to use alkylnaphthalene-sulphonates, such as diisopropyl- or diisobutylnaphthalene-sulphonates.
• Suitable dispersants and/or emulsifiers that ay be present in the seed dressing formulations include all nonionic, anionic, and cationic dispersants which are customary in the formulation of active agrochemical substances. With preference, it is possible to use nonionic or anionic dispersants or mixtures of nonionic or anionic dispersants.
• Particularly suitable nonionic dispersants are ethylene oxide- propylene oxide block polymers, alkylphenol polyglycol ethers and tristyrylphenol polyglycol ethers, and their phosphated or sulphated derivatives.
• Particularly suitable anionic dispersants are ligno sulphonates , polyacrylic salts, and arylsulphonate-formaldehyde condensates.
• Defoamers that may be present in the seed dressing formulations include all foam-inhibiting compounds which are customary in the formulation of agrochemically active compounds. Preference is given to using silicone defoamers, magnesium stearate, silicone emulsions, long-chain alcohols, fatty acids and their salts and also organofluorine compounds and mixtures thereof.
• Preservatives that may be present in the seed dressing formulation include all compounds which can be used for such purposes in agrochemical compositions. By way of example, mention may be made of dichlorophen and benzyl alcohol hemiformal.
• Secondary thickeners that may be present in the seed dressing formulations include all compounds which can be used for such purposes in agrochemical compositions. Preference is given to cellulose derivatives, acrylic acid derivatives, polysaccharides, such as xanthan gum or Veegum, modified clays, phyllosilicates, such as attapulgite and bentonite, and also finely divided silicic acids.
• Suitable adhesives that may be present in the seed dressing formulations include all customary binders which can be used in seed dressings. P oly viny lpyrrolidone , polyvinyl acetate, polyvinyl alcohol and tylose may be mentioned as being preferred.
• the gibberellins are known (Wegler "Chemie der convincedschutz- und
• the seed dressing formulations can be used directly or after dilution with water beforehand to treat seed of any of a very wide variety of types or transgenic plants. When the latter seeds are used, synergistic effects may also arise in interaction with the substances formed by expression.
• Suitable mixing equipment for treating seed with the seed dressing formulations or the preparations prepared from them by adding water includes all mixing equipment which can commonly be used for dressing.
• the specific procedure adopted when dressing comprises introducing the seed into a mixer, adding the particular desired amount of seed dressing formulation, either as it is or following dilution with water beforehand, and carrying out mixing until the formulation is uniformly distributed on the seed.
• a drying operation follows.
• the seeds are substantially uniformly coated with one or more layers of the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and/or the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant using conventional methods of mixing, spraying or a combination thereof through the use of treatment application equipment that is specifically designed and manufactured to accurately, safely, and efficiently apply seed treatment products to seeds.
• treatment application equipment uses various types of coating technology such as rotary coaters, drum coaters, fluidized bed techniques, spouted beds, rotary mists or a combination thereof.
• Liquid seed treatments such as those of the present invention can be applied via either a spinning "atomizer” disk or a spray nozzle which evenly distributes the seed treatment onto the seed as it moves though the spray pattern.
• the seed is then mixed or tumbled for an additional period of time to achieve additional treatment distribution and drying.
• the seeds can be primed or unprimed before coating with the compound (1-1 ), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant to increase the uniformity of germination and emergence.
• a dry powder formulation can be metered onto the moving seed and allowed to mix until completely distributed.
• the seeds may be coated via a batch or continuous coating process.
• continuous flow equipment simultaneously meters both the seed flow and the seed treatment products.
• a slide gate, cone and orifice, seed wheel, or weighing device (belt or diverter) regulates seed flow. Once the seed flow rate through treating equipment is determined, the flow rate of the seed treatment is calibrated to the seed flow rate in order to deliver the desired dose to the seed as it flows through the seed treating equipment. Additionally, a computer system may monitor the seed input to the coating machine, thereby maintaining a constant flow of the appropriate amount of seed.
• batch treating equipment weighs out a prescribed amount of seed and places the seed into a closed treating chamber or bowl where the corresponding dose of seed treatment is then applied. This batch is then dumped out of the treating chamber in preparation for the treatment of the next batch. With computer control systems, this batch process is automated enabling it to continuously repeat the batch treating process.
• the seed coating machinery can optionally be operated by a programmable logic controller that allows various equipments to be started and stopped without employee intervention.
• the components of this system are commercially available through several sources such as Gustafson Equipment of Shakopee, MN.
• any plant seed capable of germinating to form a plant that is susceptible to attack by insects, mites nematodes and/ or pathogenic fungi can be treated in accordance with the invention.
• Particularly suitable convential (i.e. not being a transgenic seeds) or transgenic seeds are those of cole crops, vegetables (in particular the vegetables as mentioned herein as being horticultural crops), fruits (in particular the vegetables as mentioned herein as being horticultural crops), trees, fiber crops, oil crops, tuber crops, coffee, flowers, legume, cereals, as well as other plants of the monocotyledonous and dicotyledonous species. Preference is given to seeds of horticultural crops and of broad acre crops as mentioned herein.
• seeds to be coated include soybean, cotton, corn, peanut, tobacco, grasses, wheat, barley, rye, sorghum, rice, rapeseed, sugar beet, sunflower, tomato, pepper, bean, lettuce, potato, and carrot seeds.
• the plants emerging from these seeds are capable of the expression of a protein directed against pests and pathogens.
• the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant certain pests and/or phytopathogens can already be controlled by expression of the, for example, insecticidal protein, and it is additionally surprising that a synergistic activity supplementation occurs when the compound (1-1), (1-2) or (1-3) and the biological control agents are used or employed for seed treatment, thereby improving still further the effectiveness of the protection from pest and pathogen infestation.
• Arthropoda From the class of the Arachnida, for example Acarus spp., Aceria sheldoni, Aculops spp., Aculus spp., Amblyomma spp., Amphite tranychus viennensis, Argas spp., Boophilus spp., Brevipalpus spp., Bryobia praetiosa, Centruroides spp., Chorioptes spp., Dermanyssus gallinae, Dermatophagoides pteronyssius, Dermatophagoides farinae, Dermacentor spp., Eotetranychus spp., Epitrimerus pyri, Eutetranychus spp., Eriophyes spp., Halotydeus destructor, Hemitarsonemus spp., Hyalomma spp., Ixodes spp., Lat
• Panonychus spp. Phyllocoptruta oleivora, Polyphagotarsonemus latus, Psoroptes spp., Rhipicephalus spp., Rhizoglyphus spp., Sarcoptes spp., Scorpio maurus, Stenotarsonemus spp., Tarsonemus spp., Tetranychus spp., Vaejovis spp., Vasates lycopersici.
• Anoplura for example, Damalinia spp., Haematopinus spp., Linognathus spp., Pediculus spp., Ptirus pubis, Trichodectes spp..
• Aedes spp. From the order of the Diptera, for example, Aedes spp., Agromyza spp., Anastrepha spp., Anopheles spp., Asphondylia spp., Bactrocera spp., Bibio hortulanus, Calliphora erythrocephala, Ceratitis capitata, Chironomus spp., Chrysomyia spp., Chrysops spp., Cochliomyia spp., Contarinia spp., Cordylobia anthropophaga, Culex spp., Culicoides spp., Culiseta spp., Cuterebra spp., Dacus oleae, Dasyneura spp., Delia spp., Dermatobia hominis, Drosophila spp., Echinocnemus spp., Fanni
• Anuraphis cardui, Aonidiella spp., Aphanostigma piri, Aphis spp., Arboridia apical is, Aspidiella spp., Aspidiotus spp., Atanus spp., Aidacorthum solani, Bemisia spp., Brachycaudus helichrysii, Brachycolus spp., Brevicoryne brassicae, Calligypona marginata, Carneocephala fulgida, Ceratovacuna lanigera, Cercopidae, Ceroplastes spp., Chaetosiphon fragaefolii, Chionaspis tegalensis, Chlorita onukii, Chromaphis juglandicola, Chrysomphalus ficus, Cicadulina mbila,
• Coccomytilus halli, Coccus spp., Cryptomyzus rib is, Dalbulus spp., Dialeurodes spp., Diaphorina spp., Diaspis spp., Drosicha spp., Dysaphis spp., Dysmicoccus spp., Empoasca spp., Eriosoma spp., Erythroneura spp., Euscelis bilobatus, Ferrisia spp., Geococcus coffeae, Hieroglyphus spp., Homalodisca coagulata, Hyalopterus arundinis, Icerya spp., Idiocerus spp., Idioscopus spp., Lao- delphax striatellus, Lecanium spp., Lepidosaphes spp., Lipaphis erysimi, Macrosiphum spp
• Mahanarva spp. Melanaphis sacchari, Metcalfiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzus spp., Nasonovia ribisnigri, Nephotettix spp., Nilaparvata lugens, Oncometopia spp., Orthezia praelonga, Parabemisia myricae, Paratrioza spp., Parlatoria spp., Pemphigus spp., Peregrinus maidis, Phenacoccus spp., Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp., Pinnaspis aspidistrae, Planococcus spp., Protopulvinaria pyriformis,
• Pseudaulacaspis pentagona Pseudococcus spp., Psylla spp., Pteromalus spp., Pyrilla spp., Quadraspidiotus spp., Quesada gigas, Rastrococcus spp., Rhopalosiphum spp., Saissetia spp., Scaphoides titanus, Schiz aphis graminum, Selenaspidus articulatus, Sogata spp., Sogatella furcifera, Sogatodes spp., Stictocephala festina, Tenalaphara malayensis, Tinocallis caryaefoliae, Tomaspis spp., Toxoptera spp., Trialeurodes spp., Trioza spp., Typhlocyba spp., Unaspis spp.,
• Viteus vitifolii, Zygina spp. From the order of the Hymenoptera, for example, Acromyrmex spp., Athalia spp., Atta spp., Diprion spp., Hoplocampa spp., Lasius spp., Monomorium pharaonis, Solenopsis invicta, Tapinoma spp., Vespa spp..
• Isopoda for example, Armadillidi m vulgare, Oniscus asellus, Porcellio scaber.
• Coptotermes spp. From the order of the Isoptera, for example, Coptotermes spp., Cornitermes cumulans, Cryptoterm.es spp., Incisitermes spp., Microtermes obesi, Odontotermes spp., Reticulitermes spp..
• Lepidoptera From the order of the Lepidoptera, for example, Acronicta major, Adoxophyes spp., Aedia leucomelas, Agrotis spp., Alabama spp., Amyelois transitella, Anarsia spp., Anticarsia spp., Argyroploce spp., Barathra brassicae, Borbo cinnara, Bucculatrix thurberiella, Bupalus piniarius,
• Busseola spp. Cacoecia spp., Caloptilia theivora, Capua reticulana, Carpocapsa pomonella, Carposina niponensis, Cheimatobia brumata, Chilo spp., Choristoneura spp., Clysia ambiguella, Cnaphalocerus spp., Cnephasia spp., Conopomorpha spp., Conotrachelus spp., Copitarsia spp., Cydia spp., Dalaca noctuides, Diaphania spp., Diatraea saccharalis, Earias spp., Ecdytolopha aurantium, Elasmopalpus lignosellus, Eldana saccharina, Ephestia spp., Epinotia spp., Epiphyas postvittana, Eti
• Lymantria spp. Lyonetia spp., Malacosoma neustria, Maruca testulalis, Mamestra brassicae, Mocis spp., Mythimna separata, Nymphula spp., Oiketicus spp., Oria spp., Orthaga spp., Ostrinia spp., Oulema oryzae, Panolis flammea, Parnara spp., Pectinophora spp., Perileucoptera spp., Phthorimaea spp., Phyllocnistis citrella, Phyllonorycter spp., Pieris spp., Platynota stultana, Plodia interpunctella, Plusia spp., Plutella xylostella, Prays spp., Prodenia spp., Protoparce spp.
• Pseudaletia spp., Pseudoplusia includens, Pyrausta nubilalis, Rachiplusia nu, Schoenobius spp., Scirpophaga spp., Ontario segetum, Sesamia spp., Sparganothis spp., Spodoptera spp., Stathmopoda spp., Stomopteryx subsecivella, Synanthedon spp., Tecia solanivora, Thermesia gemmatalis, Tinea pellionella, Tineola bisselliella, Tortrix spp., Trichophaga tapetzella, Trichoplusia spp., Tuta absoluta, Virachola spp..
• Orthoptera From the order of the Orthoptera, for example, Acheta domesticus, Blatta orientalis, Blattella germanica, Dichroplus spp., Gryllotalpa spp., Leucophaea maderae, Locusta spp., Melanoplus spp., Periplaneta spp., Pulex irritans, Schistocerca gregaria, Supella longipalpa.
• Siphonaptera for example, Ceratophyllus spp., Ctenocephalides spp., Tunga penetrans, Xenopsylla cheopis.
• Mollusca From the class of the Bivalvia, for example, Dreissena spp.. From the class of the Gastropoda, for example, Arion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., Galba spp., Lymnaea spp., Oncomelania spp., Pomacea spp., Succinea spp..
• Plathelminthes Nematodes (animal parasites): From the class of the Helminths, for example, Ancylostoma duodenale, Ancylostoma ceylanicum, Acylostoma braziliensis, Ancylostoma spp., Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp, Dictyocaulus filaria, Diphyl lobothrium latum, Dracunculus medinensis, Echinococcus granulosus, Echinococcus multilocularis, Enterobius vermicularis, Faciola spp., Haemonchus spp., Heterakis spp., Hymenolepis nana, Hyostrongulus
• Nematodes plant parasites, phytoparasites: From the group of the phytoparasitic nematodes, for example, Aphelenchoides spp., Bursaphelenchus spp., Ditylenchus spp., Globodera spp., Heterodera spp., Longidorus spp., Meloidogyne spp., Pratylenchus spp., Radopholus similis, Trichodorus spp., Tylenchulus semipenetrans, Xiphinema spp..
• Subphylum Protozoa such as Eimeria.
• Blumeria species such as, for example, Blumeria graminis
• Podosphaera species such as, for example, Podosphaera leuco- tricha
• Sphaerotheca species such as, for example, Sphaerotheca fuliginea
• Uncinula species such as, for example, Uncinula necator
• rust disease pathogens such as, for example, Gymnosporangium species, such as, for example, Gymnosporangium sabinae; Hemileia species, such as, for example, Hemileia vastatrix; Phakopsora species, such as, for example, Phakopsora pachyrhizi and Phakopsora meibomiae; Puccinia species, such as, for example, Puccinia recondita or Puccinia triticina; Uromyces species, such as, for example, Uromyces appendiculatus; Diseases caused by pathogens from the group of the Oomycetes, such as, for example, Albugo species, such as, for example, Albugo Candida; Bremia species, such as, for example, Bremia lactucae; Peronospora species, such as, for example, Peronospora pisi or P.
• Gymnosporangium species such as, for example, Gymnospor
• Phytophthora species such as, for example Phytophthora infestans
• Plasmopara species such as, for example, Plasmopara viticola
• Pseudoperonospora species such as, for example,
• Pseudoperonospora hamuli or Pseudoperonospora cubensis Pythium species, such as, for example, Pythium ultimum;
• Venturia species such as, for example, Venturia inaequalis
• Root sheath and stem diseases caused, for example, by Corticium species, such as, for example, Corticium graminearum; Fusarium species, such as, for example, Fusarium oxysporum;
• Gaeumannomyces species such as, for example, Gaeumannomyces graminis; Rhizoctonia species, such as, for example Rhizoctonia solani; Sarocladium species, such as, for example, Sarocladium oryzae; Sclerotium species, such as, for example, Sclerotium oryzae; Tapesia species, such as, for example, Tapesia acuformis; Thielaviopsis species, such as, for example, Thielaviopsis basicola; Ear and panicle diseases (including maize cobs) caused, for example, by Alternaria species, such as, for example, Alternaria spp.; Aspergillus species, such as, for example, Aspergillus flavus; Cladosporium species, such as, for example, Cladosporium cladosporioides; Claviceps species, such as, for example, Claviceps purpurea; Fusarium species, such as, for example
• Pyrenophora diseases caused for example by Pyrenophora graminea; Pyricularia diseases caused for example by Pyricularia oryzae; Pythium diseases caused for example by Pythium ultimum; Rhizoctonia diseases caused for example by Rhizoctonia solani; Rhizopus diseases caused for example by Rhizopus oryzae; Sclerotium diseases caused for example by Sclerotium rolfsii; Septoria diseases caused for example by Septoria nodorum; Typhula diseases caused for example by Typhula incarnata; Verticillium diseases caused for example by Verticillium dahliae;
• Sphacelotheca species such as, for example, Sphacelotheca reiliana
• Tilletia species such as, for example, Tilletia caries
• Tilletia controversa Tilletia species
• Urocystis species such as, for example, Urocystis occulta
• Ustilago species such as, for example, Ustilago nuda
• Aspergillus species such as, for example, Aspergillus flavus
• Botiytis species such as, for example, Botrytis cinerea
• Penicillium species such as, for example, Penicillium expansum and Penicillium purpurogenum
• Rhizopus species such as, for example, Rhizopus stolonifer
• Sclerotinia species such as, for example
• Typhula species such as, for example, Typhula incarnate
• VerticiUium species such as, for example, VerticiUium dahliae
• Nectria species such as, for example, Nectria galligena
• Exobasidium species such as, for example, exobasidium vexans
• Taphrina species such as, for example, Taphrina deformans
• Phaemoniella clamydospora and Phaeoacremonium aleophilum and Fomitiporia mediterranea Ganoderma species, such as, for example, Ganoderma boninense; Rigidoporus species, such as, for example, Rigidoporus lignosus;
• Plasmodiophora species such as, for example, Plasmodiophora brassicae
• Botrytis species such as, for example, Botrytis cinerea
• Rhizoctonia species such as, for example, Rhizoctonia solani
• Helminthosporium species such as, for example, Helminthosporium solani
• Diseases caused by bactenopathogens such as, for example, Xanthomonas species, such as, for example, Xanthomonas campestris pv. oryzae
• Pseudomonas species such as, for example, Pseudomonas syringae pv. lachrymans
• Erwinia species such as, for example, Erwinia amylovora.
• phytophthora rot (Phytophthora megasperma), brown stem rot (Phialophora gregata), pythium rot (Pythium aphanidermatum, Pythium irregulare, Pythium debaryanum, Pythium myriotylum, Pythium ultimum), rhizoctonia root rot, stem decay, and damping-off (Rhizoctonia solani), sclerotinia stem decay (Sclerotinia sclerotiorum), sclerotinia Southern blight
• the compound (1-1), (1-2) or (1-3) in combination with at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant as given herein can also exhibit very good antimycotic activity, in particular against dermatophytes and yeasts, moulds and diphasic fungi (for example against Candida species such as Candida albicans, Candida glabrata) and
• Epidermophyton floccosum Aspergillus species such as Aspergillus niger and Aspergillus fumigatus, Trichophyton species such as Trichophyton mentagrophytes, Microsporon species such as Microsporon canis and audouinii.
• the list of these fungi by no means limits the mycotic spectrum which can be covered, but is only for illustration.
• the compound (1-1), (1-2) or (1-3) in combination with at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant as given herein can also be used to reduce the contents of mycotoxins in plants and the harvested plant material and therefore in foods and animal feed stuff made therefrom.
• at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant as given herein can also be used to reduce the contents of mycotoxins in plants and the harvested plant material and therefore in foods and animal feed stuff made therefrom.
• mycotoxins can be specified: Deoxynivalenole (DON), Nivalenole, 15-Ac-DON, 3-Ac-DON, T2- und I IT2- Toxins, Fumonisines, Zearalenone Moniliformine, Fusarine, Diaceotoxyscirpenole (DAS), Beauvericine, Enniatine, Fusaroproliferine, Fusarenole, Ochratoxines, Patuline, Ergotalkaloides und Aflatoxines, which are caused for example by the following fungal diseases: Fusarium spec, like Fusarium acuminatum, F. avenaceum, F. crookwellense, F.
• plants and plant parts which are mentioned herein are all plants and plant populations, such as desired and undesired wild plants or crop plants (including naturally occurring crop plants).
• Crop plants can be plants which can be obtained by traditional breeding and optimization methods or by biotechnological and recombinant methods, or combinations of these methods, including the transgenic plants and including the plant varieties capable or not of being protected by Plant Breeders' Rights.
• Plant parts are understood as meaning all aerial and subterranean parts and organs of the plants, such as shoot, leaf, flower and root, examples which may be mentioned being leaves, needles, stalks, stems, flowers, fruiting bodies, fruits and seeds, and also roots, tubers and rhizomes.
• the plant parts also include crop material and vegetative and generative propagation material, for example cuttings, tubers, rhizomes, slips and seeds.
• plants and their parts ay be treated in accordance with the invention.
• plant species and plant varieties, and their parts which grow wild or which are obtained by traditional biological breeding methods such as hybridization or protoplast fusion are treated.
• transgenic plants and plant varieties which have been obtained by recombinant methods, i appropriate in combination with traditional methods (genetically modified organisms), and their parts are treated.
• the term "parts” or “parts of plants” or “plant parts” has been explained hereinabove. Plants of the plant varieties which are in each case commercially available or in use are especially preferably treated in accordance with the invention. Plant varieties are understood as meaning plants with novel traits which have been bred both by traditional breeding, by mutagenesis or by recombinant D A techniques. The may take the form of varieties, races, biotypes and genotypes.
• Preferred plants are those from the group of the useful plants, ornamentals, turfs, generally used trees which are employed as ornamentals in the public and domestic sectors, and forestry trees.
• Forestry trees comprise trees for the production of timber, cellulose, paper and products made from parts of the trees.
• useful plants refers to crop plants which are employed as plants for obtaining foodstuffs, feedstuffs, fuels or for industrial purposes.
• compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant include for example the following types of plants: turf, vines, cereals, for example wheat, barley, rye, oats, rice, maize and millet/sorghum; beet, for example sugar beet and fodder beet; fruits, for example pome fruit, stone fruit and soft fruit, for example apples, pears, plums, peaches, almonds, cherries and berries, for example strawberries, raspberries, blackberries;
• legumes for example beans, lentils, peas and soybeans
• oil crops for example oilseed rape, mustard, poppies, olives, sunflowers, coconuts, castor oil plants, cacao and peanuts
• cucurbits for example pumpkin/squash, cucumbers and melons
• fibre plants for example cotton, flax, hemp and jute
• citrus fruit for example oranges, lemons, grapefruit and tangerines
• vegetables for example spinach, lettuce, asparagus, cabbage species, carrots, onions, tomatoes, potatoes and bell peppers
• Lauraceae for example avocado, Cinnamomum, camphor, or else plants such as tobacco, nuts, coffee, aubergine, sugar cane, tea, pepper, grapevines, hops, bananas, latex plants and ornamentals, for example flowers, shrubs, deciduous trees and coniferous trees.
• the following plants are considered to be particularly suitable target crops for using or employing the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inocuiant: cotton, aubergine, turf, pome fruit, stone fruit, soft fruit, maize, wheat, barley, cucumber, tobacco, vines, rice, cereals, pear, beans, soybeans, oilseed rape, tomato, bell pepper, melons, cabbage, potato and apple.
• bacteria in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes
• the inocuiant cotton, aubergine, turf, pome fruit, stone fruit, soft fruit, maize, wheat, barley, cucumber, tobacco, vines, rice, cereals, pear, beans, soybean
• Examples of trees are: Abies sp., Eucalyptus sp., Picea sp., Pinus sp., Aesculus sp., Platanus sp., Tilia sp., Acer sp., Tsuga sp., Fraxinus sp., Sorbus sp., Betula sp., Crataegus sp., Ulmus sp., Quercus sp., Fagus sp., Salix sp., Populus sp..
• Preferred trees which can be improved in accordance with the method according to the invention are: from the tree species Aesculus: A. hippocastanum, A. pariflora, A. cornea; from the tree species Platanus: P. aceriflora, P. occidentalis, P. racemosa; from the tree species Picea: P. abies; from the tree species Pinus: P. radiata, P. ponderosa, P. contorta, P. sylvestre, P. elliottii, P. montecola, P. albicaulis, P. resinosa, P. palustris, P. taeda, P. flexilis, P. jeffregi, P. baksiana, P. strobus; from the tree species Eucalyptus: E. grandis, E. globulus, E. camadentis, E. nitens, E. obliqua, E. regnans, E.
• Especially preferred trees which can be improved in accordance with the method according to the invention are: from the tree species Pinus: P. radiata, P. ponderosa, P. contorta, P. sylvestre, P. strobus; from the tree species Eucalyptus: E. grandis, E. globulus, E. camadentis.
• Very particularly preferred trees which can be improved in accordance with the method according to the invention are: horse chestnut, Platanaceae, linden tree, maple tree.
• the present invention can also be applied to any turf grasses, including cool-season tur grasses and warm-season turf grasses.
• cold-season turf grasses are bluegrasses (Poa spp.), such as Kentucky bluegrass (Poa pratensis L.), rough bluegrass (Poa trivialis L.), Canada bluegrass (Poa compressa L.), annual bluegrass (Poa annua L.), upland bluegrass (Poa glaucantha Gaudin), wood bluegrass (Poa nemoralis L.) and bulbous bluegrass (Poa bulbosa L.); bentgrasses (Agrostis spp.) such as creeping bentgrass (Agrostis palustris Huds.), colonial bentgrass (Agrostis tenuis Sibth.), velvet bentgrass (Agrostis canina L.), South German mixed bentgrass (Agrostis spp.
• ryegrasses such as annual ryegrass (Lolium multiflorum Lam.), perennial ryegrass (Lolium perenne L.) and Italian ryegrass (Lolium multiflorum Lam.); and wheatgrasses (Agropyron spp.), such as fairway wheatgrass (Agropyron cristatum (L.) Gaertn.), crested wheatgrass (Agropyron desertorum (Fisch.) Schult.
• Examples of further cool-season turf grasses are beachgrass (Ammophila breviligulata Fern.), smooth bromegrass (Bromus inermis Leyss.), cattails such as timothy (Phleum pratense L.), sand cattail (Phleum subulatum L.), orchardgrass (Dactylis glomerata L.), weeping alkaligrass (Puccinellia distans (L.) Pari.) and crested dog's-tail (Cynosurus cristatus L.).
• beachgrass Adophila breviligulata Fern.
• smooth bromegrass Bromus inermis Leyss.
• cattails such as timothy (Phleum pratense L.), sand cattail (Phleum subulatum L.), orchardgrass (Dactylis glomerata L.), weeping alkaligrass (Puccinellia distans (
• Examples of warm-season turf grasses are Bermuda grass (Cynodon spp. 1 C. Rich), zoysia grass (Zoysia spp. Willd.), St. Augustine grass (Stenotaphrum secundatum Walt Kuntze), centipede grass (Eremochloa ophiuroides Munrohack.), carpetgrass (Axonopus affinis Chase), Bahia grass (Paspalum notatum Flugge), Kikuyu grass (Pennisetum clandestinum Hochst.
• Cool-season turf grasses are generally preferred for the use according to the invention. Especially preferred are bluegrass, benchgrass and redtop, fescues and ryegrasses. Bentgrass is especially preferred.
• heterologous gene essentially means a gene which is provided or assembled outside the plant and when introduced in the nuclear, chloroplastic or mitochondrial genome gives the transformed plant new or improved agronomic or other properties by expressing a protein or polypeptide of interest or by down regulating or silencing other gene(s) which are present in the plant (using for example, antisense technology, co-suppression technology or RNA interference - RNAi - technology).
• a heterologous gene that is located in the genome is also called a transgene.
• a transgene that is defined by its particular location in the plant genome is called a transformation or transgenic event.
• the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore - forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant in the treatment according to the invention reduced application rates and/or a widening of the activity spectrum and/or an increase in the activity better plant growth, increased tolerance to high or low temperatures, increased tolerance to drought or to water or soil salt content, increased flowering performance, easier harvesting, accelerated maturation, higher harvest yields, bigger fruits, larger plant height, greener leaf color, earlier flowering, higher quality and/or a higher nutritional value of the harvested products, higher sugar concentration within the fruits, better storage stability and/or processability of the harvested products are possible, which exceed the effects which were actually to be expected.
• bacteria in particular spore - forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nem
• At certain application rates of the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore-forming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant in the treatment according to the invention may also have a strengthening effect in plants.
• the defense system of the plant against attack by unwanted phytopathogenic fungi and/ or microorganisms and/or viruses is mobilized.
• Plant-strengthening (resistance-inducing) substances are to be understood as meaning, in the present context, those substances or combinations of substances which are capable of stimulating the defense system of plants in such a way that, when subsequently inoculated with unwanted
• the treated plants display a substantial degree of resistance to these phytopathogenic fungi and/or microorganisms and/or viruses
• the compound (1-1), (1-2) or (1-3), preferably compound (1-3) and the at least one biological control agent selected from bacteria, in particular spore- orming bacteria, fungi or yeasts, protozoas, viruses, and entomopathogenic nematodes, and optionally the inoculant in the treatment according to the invention plants can be protected against attack by the abovementioned pathogens within a certain period of time after the treatment.
• the period of time within which protection is effected generally extends from 1 to 10 days, preferably 1 to 7 days, after the treatment of the plants with the active compounds.
• Plants and plant cultivars which are also preferably to be treated according to the invention are resistant against one or more biotic stresses, i.e. said plants show a better defense against animal and microbial pests, such as against nematodes, insects, mites, phytopathogenic fungi, bacteria, viruses and/or viroids.
• Plants and plant cultivars which may also be treated according to the invention are those plants which are resistant to one or more abiotic stresses.
• Abiotic stress conditions may include, for example, drought, cold temperature exposure, heat exposure, osmotic stress, flooding, increased soil salinity, increased mineral exposure, ozon exposure, high light exposure, limited availability of nitrogen nutrients, limited availability of phosphorus nutrients, shade avoidance.
• Plants and plant cultivars which may also be treated according to the invention are those plants characterized by enhanced yield characteristics. Increased yield in said plants can be the result of, for example, improved plant physiology, growth and development, such as water use efficiency, water retention efficiency, improved nitrogen use, enhanced carbon assimilation, improved photosynthesis, increased germination efficiency and accelerated maturation.
• Yield can furthermore be affected by improved plant architecture (under stress and non-stress conditions), including but not limited to, early flowering, flowering control for hybrid seed production, seedling vigor, plant size, internode number and distance, root growth, seed size, fruit size, pod size, pod or ear number, seed number per pod or ear, seed mass, enhanced seed filling, reduced seed dispersal, reduced pod dehiscence and lodging resistance.
• Further yield traits include seed composition, such as carbohydrate content, protein content, oil content and composition, nutritional value, reduction in anti-nutritional compounds, improved processability and better storage stability.
• Plants that may be treated according to the invention are hybrid plants that already express the characteristic of heterosis or hybrid vigor which results in generally higher yield, vigor, health and resistance towards biotic and abiotic stress factors. Such plants are typically made by crossing an inbred male-sterile parent line (the female parent) with another inbred male-fertile parent line (the male parent). I lybrid seed is typically harvested from the male sterile plants and sold to growers. Male sterile plants can sometimes (e.g. in corn) be produced by detasseling, i.e. the mechanical removal of the male reproductive organs (or males flowers) but, more typically, male sterility is the result of genetic determinants in the plant genome.
• male sterile plants can also be obtained by plant biotechnology methods such as genetic engineering.
• a particularly useful means of obtaining male-sterile plants is described in WO 89/10396 in which, for example, a ribonuclease such as bamase is selectively expressed in the tapetum cells in the stamens. Fertility can then be restored by expression in the tapetum cells of a ribonuclease inhibitor such as barstar.
• Plants or plant cultivars obtained by plant biotechnology methods such as genetic engineering which may be treated according to the invention are herbicide-tolerant plants, i.e. plants made tolerant to one or more given herbicides. Such plants can be obtained either by genetic transformation, or by selection of plants containing a mutation imparting such herbicide tolerance.
• Herbicide-tolerant plants are for example glyphosate-tolerant plants, i.e. plants made tolerant to the herbicide glyphosate or salts thereof. Plants can be made tolerant to glyphosate through different means. For example, glyphosate-tolerant plants can be obtained by transforming the plant with a gene encoding the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS).
• EPSPS 5-enolpyruvylshikimate-3-phosphate synthase
• EPSPS genes are the AroA gene (mutant CT7) of the bacterium Salmonella typhimurium, the CP4 gene of the bacterium Agrobacterium sp, the genes encoding a Petunia EPSPS, a Tomato EPSPS, or an Eleusine EPSPS. It can also be a mutated EPSPS.
• Glyphosate-tolerant plants can also be obtained by expressing a gene that encodes a glyphosate oxido-reductase enzyme.
• Glyphosate-tolerant plants can also be obtained by expressing a gene that encodes a glyphosate acetyl transferase enzyme.
• Glyphosate-tolerant plants can also be obtained by selecting plants containing naturally-occurring mutations of the above-mentioned genes.
• herbicide resistant plants are for example plants that are made tolerant to herbicides inhibiting the enzyme glutamine synthase, such as bialaphos, phosphinothricin or glufosinate.
• Such plants can be obtained by expressing an enzyme detoxifying the herbicide or a mutant glutamine synthase enzyme that is resistant to inhibition.
• One such efficient detoxifying enzyme is an enzyme encoding a phosphinothricin acetyltransferase (such as the bar or pat protein from Streptomyces species). Plants expressing an exogenous phosphinothricin acetyltransferase are also described.
• Further herbicide -tolerant plants are also plants that are made tolerant to the herbicides inhibiting the enzyme hydroxyphenylpyruvatedioxygenase (HPPD).
• HPPD hydroxyphenylpyruvatedioxygenase
• Hydroxyphenylpyruvatedioxygenases are enzymes that catalyze the reaction in which para- hydroxyphenylpyruvate (HPP) is transformed into homogentisate.
• Plants tolerant to HPPD -inhibitors can be transformed with a gene encoding a naturally-occurring resistant HPPD enzyme, or a gene encoding a mutated HPPD enzyme.
• Tolerance to HPPD-inhibitors can also be obtained by transforming plants with genes encoding certain enzymes enabling the formation of homogentisate despite the inhibition of the native HPPD enzyme by the HPPD-inhibitor.
• Tolerance of plants to HPPD inhibitors can also be improved by transforming plants with a gene encoding an enzyme prephenate dehydrogenase in addition to a gene encoding an HPPD-tolerant enzyme.
• Still further herbicide resistant plants are plants that are made tolerant to acetolactate synthase (ALS) inhibitors.
• ALS-inhibitors include, for example, sulfonylurea, imidazolinone, triazolopyrimidines, pyrimidinyoxy(thio)benzoates, and/or sulfonylaminocarbonyltriazolinone herbicides.
• Different mutations in the ALS enzyme also known as acetohydroxyacid synthase, AHAS
• AHAS acetohydroxyacid synthase
• the production of sulfonylurea-tolerant plants and imidazolinone-tolerant plants is described in WO 1996/033270. Other imidazolinone-tolerant plants are also described. Further sulfonylurea- and imidazolinone-tolerant plants are also described in for example WO 2007/024782.
• plants tolerant to imidazolinone and/or sulfonylurea can be obtained by induced mutagenesis, selection in cell cultures in the presence o the herbicide or mutation breeding as described for example for soybeans, for rice, for sugar beet, for lettuce, or for sunflower.
• Plants or plant cultivars obtained by plant biotechnology methods such as genetic engineering which may also be treated according to the invention are insect-resistant transgenic plants, i.e. plants made resistant to attack by certain target insects. Such plants can be obtained by genetic transformation, or by selection of plants containing a mutation imparting such insect resistance.
• An "insect-resistant transgenic plant”, as used herein, includes any plant containing at least one transgene comprising a coding sequence encoding;
• insecticidal crystal protein from Bacillus thuringiensis or an insecticidal portion thereof, such as the insecticidal crystal proteins listed online at:
• a crystal protein from Bacillus thuringiensis or a portion thereof which is insecticidal in the presence of a second other crystal protein from Bacillus thuringiensis or a portion thereof, such as the binary toxin made up of the Cry 34 and Cry 35 crystal proteins; or
• a hybrid insecticidal protein comprising parts of different insecticidal crystal proteins from Bacillus thuringiensis, such as a hybrid of the proteins o 1) above or a hybrid of the proteins of 2) above, e.g., the CrylA.105 protein produced by corn event MON98034 (WO 2007/027777); or
• VIP vegetative insecticidal
• proteins from the VIP3Aa protein class e.g. proteins from the VIP3Aa protein class; or 6) secreted protein from Bacillus thuringiensis or Bacillus cereus which is insecticidal in the presence of a second secreted protein from Bacillus thuringiensis or B. cereus, such as the binary toxin made up of the VIP1 A and VIP2A proteins; or
• hybrid insecticidal protein comprising parts from different secreted proteins from Bacillus thuringiensis or Bacillus cereus, such as a hybrid of the proteins in 1) above or a hybrid of the proteins in 2) above; or
• Plants or plant cultivars obtained by plant biotechnology methods such as genetic engineering
• plants or plant cultivars which may also be treated according to the invention are nematode-resistant transgenic plants, i.e. plants made resistant to attack by certain target nematode.
• Such plants can be obtained by genetic transformation, or by selection of plants containing a mutation imparting such insect resistance
• Examples of nematode resistant plants are described in e.g. US Patent Application os 11/765,491, 11/765,494, 10/926,819, 10/782,020, 12/032,479, 10/783,417, 10/782,096, 11/657,964, 12/192,904, 11/396,808, 12/166,253, 12/166,239, 12/166,124, 12/166,209, 11/762,886, 12/364,335, 11/763,947, 12/252,453, 12/209,354, 12/491,396 or 12/497,221.
• an insect-resistant transgenic plant also includes any plant comprising a combination of genes encoding the proteins of any one of the above classes 1 to 8.
• an insect-resistant plant contains more than one transgene encoding a protein of any one of the above classes 1 to 8, to expand the range of target insect species affected when using different proteins directed at different target insect species, or to delay insect resistance development to the plants by using different proteins insecticidal to the same target insect species but having a different mode of action, such as binding to different receptor binding sites in the insect.
• Plants or plant cultivars obtained by plant biotechnology methods such as genetic engineering which may also be treated according to the invention are tolerant to abiotic stresses. Such plants can be obtained by genetic transformation, or by selection of plants containing a mutation imparting such stress resistance. Particularly useful stress tolerance plants include:
• plants which contain a transgene capable of reducing the expression and/or the activity of p oly ( ADP -ribo s e)p oly meras e (PARP) gene in the plant cells or plants
• Plants or plant cultivars obtained by plant biotechnology methods such as genetic engineering which may also be treated according to the invention show altered quantity, quality and/or storage-stability of the harvested product and/or altered properties of specific ingredients of the harvested product such as :
• transgenic plants which synthesize a modified starch, which in its physical-chemical characteristics, in particular the amylose content or the amylose/amylopectin ratio, the degree of branching, the average chain length, the side chain distribution, the viscosity behaviour, the gelling strength, the starch grain size and/or the starch grain morphology, is changed in comparison with the synthesised starch in wild type plant cells or plants, so that this is better suited for special applications,
• transgenic plants which synthesize non starch carbohydrate polymers or which synthesize non starch carbohydrate polymers with altered properties in comparison to wild type plants without genetic modification.
• Examples are plants producing polyfructose, especially of the inulin and levan-type, plants producing alpha 1,4 glucans, plants producing alpha- 1,6 branched alpha- 1,4- glucans, plants producing alternan,
• Plants or plant cultivars that can be obtained by plant biotechnology methods such as genetic engineering
• plants which may also be treated according to the invention are plants, such as cotton plants, with altered fiber characteristics.
• Such plants can be obtained by genetic transformation or by selection of plants contain a mutation imparting such altered fiber characteristics and include:
• Plants such as cotton plants, containing an altered form of cellulose synthase genes
• Plants such as cotton plants, containing an altered form of rsw2 or rsw3 homologous nucleic acids
• Plants such as cotton plants, wherein the timing of the plasmodesmatal gating at the basis of the fiber cell is altered, e.g. through downregulation of fiberselective ⁇ 1,3-glucanase
• Plants such as cotton plants, having fibers with altered reactivity, e.g. through the expression of N-acteylglucosaminetransferase gene including nodC and chitinsynthase genes.
• Plants or plant cultivars that can be obtained by plant biotechnology methods such as genetic engineering
• plants which may also be treated according to the invention are plants, such as oilseed rape or related Brassica plants, with altered oil profile characteristics.
• Such plants can be obtained by genetic transformation or by selection of plants contain a mutation imparting such altered oil characteristics and include:
• transgenic plants which may be treated according to the invention are plants which comprise one or more genes which encode one or more toxins, such as the following which are sold under the trade names YIELD GARD® (for example maize, cotton, soya beans), KnockOut® (for example maize), BiteGard® (for example maize), Bt-Xtra® (for example maize), StarLink® (for example maize), Bollgard® (cotton), ucotn® (cotton), Nucotn 33 ⁇ ⁇ (cotton), atureGard® (for example maize), Protecta® and ewLeaf® (potato).
• YIELD GARD® for example maize, cotton, soya beans
• KnockOut® for example maize
• BiteGard® for example maize
• Bt-Xtra® for example maize
• StarLink® for example maize
• Bollgard® cotton
• ucotn® cotton
• Nucotn 33 ⁇ ⁇ cotton
• atureGard® for example
• herbicide-tolerant plants examples include maize varieties, cotton varieties and soya bean varieties which are sold under the trade names Roundup Ready® (tolerance to glyphosate, for example maize, cotton, soya bean), Liberty Link® (tolerance to phosphinotricin, for example oilseed rape), IMI® (tolerance to imidazolinones) and STS® (tolerance to sulphonylureas, for example maize).
• Herbicide-resistant plants plants bred in a conventional manner for herbicide tolerance
• Clearfield® for example maize.
• transgenic plants which may be treated according to the invention are plants containing transformation events, or a combination of transformation events, and that are listed for example in the databases for various national or regional regulatory agencies including Event 1143-14A (cotton, insect control, not deposited, described in WO 06/128569); Event 1 143- 1 B (cotton, insect control, not deposited, described in WO 06/128570); Event !
• Event 445 (cotton, herbicide tolerance, not deposited, described in US-A 2002-120964 or WO 02/034946); Event 17053 (rice, herbicide tolerance, deposited as PTA-9843, described in WO 10/117737); Event 17314 (rice, herbicide tolerance, deposited as PTA-9844, described in WO 10/117735); Event 281-24-236 (cotton, insect control - herbicide tolerance, deposited as PTA-6233, described in WO 05/103266 or US-A 2005-216969); Event 3006-210- 23 (cotton, insect control - herbicide tolerance, deposited as PTA-6233, described in US-A 2007-143876 or WO 05/103266); Event 3272 (corn, quality trait, deposited as PTA-9972, described in WO 06/098952 or US-A 2006-230473); Event 40416 (corn, insect control - herbicide tolerance, deposited as ATCC ⁇ - 1 1508, described in WO 1 1/075593); Event 43
• Event CE43-67B (cotton, insect control, deposited as DSM ACC2724, described in US 2009-217423 or WO 06/128573); Event CE44-69D (cotton, insect control, not deposited, described in US 2010-0024077); Event CE44-69D (cotton, insect control, not deposited, described in WO 06/128571); Event CE46-02A (cotton, insect control, not deposited, described in WO 06/128572); Event COT102 (cotton, insect control, not deposited, described in US 2006-130175 or WO 04/039986); Event COT202 (cotton, insect control, not deposited, described in US 2007-067868 or WO 05/054479); Event COT203 (cotton, insect control, not deposited, described in WO 05/054480); Event DAS40278 (corn, herbicide tolerance, deposited as ATCC PTA-10244, described in WO 1 1/022469); Event
• Event GJl 1 (corn, herbicide tolerance, deposited as ATCC 209030, described in US 2005-188434 or WO 98/044140); Event GM RZ13 (sugar beet, virus resistance , deposited as NCIMB-41601, described in WO 10/076212); Event 1 17- 1 (sugar beet, herbicide tolerance, deposited as NCIMB 41 3 58 or NCIMB 41 159, described in US 2004-172669 or WO 04/074492); Event JOPLINl (wheat, disease tolerance, not deposited, described in US 2008-064032); Event LL27 (soybean, herbicide tolerance, deposited as NCIMB41658, described in WO 06/108674 or US 2008-320616); Event 1.1 55 (soybean, herbicide tolerance, deposited as NCIMB 41660, described in WO 06/108675 or US 2008-196127); Event LLcotton25 (cotton, herbicide tolerance, deposited as ATCC PTA-3343,
• Event MO 89788 (soybean, herbicide tolerance, deposited as ATCC PTA-6708, described in US-A 2006-282915 or WO 06/130436); Event MSI 1 (oilseed rape, pollination control - herbicide tolerance, deposited as ATCC PTA-850 or PTA-2485, described in WO 01/031042); Event MS8 (oilseed rape, pollination control - herbicide tolerance, deposited as ATCC PTA-730, described in WO 01/041558 or US-A 2003-188347); Event K603 (corn, herbicide tolerance, deposited as ATCC PTA-2478, described in US-A 2007-292854); Event PE-7 (rice, insect control, not deposited, described in WO 08/114282); Event RF3 (oilseed rape, pollination control - herbicide tolerance, deposited as ATCC PTA-730, described in WO 01/041558 or US 2003-1883
• transgenic plants which may be treated according to the invention are plants containing transformation events, or combination of transformation events, that are listed for example in the databases from various national or regional regulatory agencies (see for example
• X is the efficacy expressed in % mortality of the untreated control for test compound A at a concentration of m ppm or m g/ha,
• Y is the efficacy expressed in % mortality of the untreated control for test compound B at a concentration o n ppm or m g/ha,
• E is the efficacy expressed in % mortality of the untreated control using the combination of A and B at m and n ppm or m and n g/ha,
• Example 1 Myzus persicae - foliar application
• Emulsifier 2 parts by weight of alkylaryl polyglycolether
• Cabbage leaves (Brassica oleracea) which are heavily infested by the Green peach aphid (Myzus persicae) are treated by being sprayed with the preparation of the active compound at the desired concentration.
• mortality in % is determined. 100 % means that all the aphids have been killed; 0 % means that none of the aphids have been killed. The mortality values thus determined are recalculated using the Colby-formula.
• Emulsifier parts by weight alkylarylpolyglycolether
• Emulsifier 2 parts by weight of alkylaryl polyglycolether
• Cabbage leaves (Brassica oleracea) are treated by being sprayed with the preparation of the active compound of the desired concentration and are infested with larvae of the mustard beetle (Phaedon cochleariae) as long as the leaves are still moist.
• Emulsifier 0.5 parts by weight of alkylaryl polyglycolether
• mortality in % is determined. 100 % means that all the beetle larvae have been killed; 0 % means that none o the beetle larvae have been killed. The mortality values thus determined are recalculated using the Colby-formula.
• Emulsifier 2 parts by weight of alkylaryl polyglycolether [0240] To produce a suitable preparation of active compound, 1 part by weight of active compound is mixed with the stated amount of solvent and emulsifier, and the concentrate is diluted with emulsifier- containing water to the desired concentration. To produce a suitable preparation of a spore suspension the spores are diluted with emulsifier- containing water to the desired concentration.
• the preparation containing the active ingredient is thoroughly mixed into the soil.
• the treated soil is filled into pots and sown with 5 corn seeds per pot. 3 days after sowing larvae of the Banded Cucumber Beetle ( Diabrotica baltcata) are placed in the soil.
• the level of activity expressed in % is determined.
• the level of activity is calculated on the basis of the number of corn plants which have successfully germinated.
• the efficacy values determined thus are recalculated using the Colby-formula.
• Seed treatment- germination test soybean [0244] Seeds of soybean (Glycine max) were treated by being mixed with the desired amount of active compound and spores and water. After drying, 25 seeds were sown into each pot filled with sandy loam.

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